Vaccines against herpes simplex virus type 2: compositions and methods for eliciting an immune response

ABSTRACT

Herpes Simplex Virus-2 (HSV-2) infection is a major health concern. The present disclosure provides, inter alia, certain highly effective vaccines and immunogenic compositions against HSV-2. The antigens can be used therapeutically or prophylactically.

RELATED APPLICATIONS

This application is a division of U.S. application Ser. No. 14/077,676,filed Nov. 12, 2013, now U.S. Pat. No. 9,895,436, which is acontinuation of U.S. application Ser. No. 12/786,425, filed May 24,2010, now U.S. Pat. No. 8,617,564, which claims the benefit of U.S.Provisional Application No. 61/180,784, filed on May 22, 2009, U.S.Provisional Application No. 61/235,628, filed on Aug. 20, 2009, U.S.Provisional Application No. 61/240,587, filed on Sep. 8, 2009, U.S.Provisional Application No. 61/240,626, filed on Sep. 8, 2009, and U.S.Provisional Application No. 61/305,918 filed on Feb. 18, 2010, theentire contents of each of which are incorporated herein by reference.

I. BACKGROUND

Herpes simplex virus type 2 (HSV-2) is the leading cause of genitalherpes. HSV-2 is most often transmitted by sexual contact, and infectionwith the virus typically leads to recurring outbreaks of lesions on thegenitals and perianal regions, combined with shedding of virus particlesinto the genital tract. Viral shedding can also occur in the absence oflesions or other symptoms. HSV-2 also establishes latency in sensoryganglia. HSV-2 infection causes physical discomfort and psychosexualmorbidity in affected patients, and introduces additional health risks.In particular, patients infected with HSV-2 are at increased risk forcontracting HIV, and pregnant mothers infected with HSV-2 can verticallytransmit HSV-2 to their fetuses. In immunocompromised individuals or inneonates, HSV-2 infections can be fatal. Currently, there is no cure forHSV-2 infection.

HSV-2 infection is widespread, with one study estimating that nearly 20%of the population worldwide is infected (Looker et al., 2008, Bulletinof the World Health Organization, October 2008, 86(10)). More women thanmen are infected, and the prevalence of the disease increases with age.High numbers of adolescents diagnosed with HSV-2 indicate that theprevalence across the population will continue to rise, as HSV-2infection is lifelong.

Treatment options for HSV-2 symptoms are limited. Antiviral therapy,using compounds such as famciclovir, valaciclovir, or aciclovir, limitsthe duration of symptoms and, in some cases, speeds healing of lesionsand reduces incidence of viral shedding. Antiviral drugs are notcurative, however, and do not prevent recurrence of outbreaks or clearthe virus completely. In addition, use of antiviral drugs requirespatients to recognize symptoms of HSV-2 infection, then obtain aconfirmative diagnosis, and ultimately, comply with the antiviralregimen. These requirements may be untenable in regions of the worldwhere antiviral drugs are not readily available. In addition, patientsare often unaware that they are infected, either because they do notpresent symptoms, or because the symptoms of the initial infectionsubside, suggesting recovery from the disease.

To address the medical and social problems associated with HSV-2, it ishighly desirable to develop pharmaceutical compositions to inhibit orcounteract infection by HSV-2. An effective composition may be used toelicit an enhanced immune response against HSV-2, thereby preventinginitial infection, blocking the ability of the virus to establishlatency in sensory ganglia, eliminating recurrence of outbreaks, and/orpreventing viral shedding. The immune system is known to mount a defenseagainst HSV-2, as evidenced by recurrent infections which manifest withfewer, less intense symptoms and decreased frequency over time.

While the ultimate goal of an HSV vaccine would be long-lastingprotection from viral infection, the suppression of disease symptomswould also provide significant health benefits. One of the current goalsfor either a prophylactic or therapeutic vaccine is to reduce clinicalepisodes and viral shedding from primary and latent infections. Threecategories of prophylactic vaccines have been tested in clinical trialswith disappointing results i) whole virus, ii) protein subunit and iii)gene-based subunit vaccines (Stanberry et al., Clinical Infect. Dis.,30(3):549-566, 2000). In the 1970s a number of killed virus vaccineswere explored, none of which were efficacious. More recently anattenuated HSV was found to be poorly immunogenic. Subunit vaccinesbased on two recombinant glycoproteins have been clinically evaluated incombination with different adjuvant formulations. One developed byChiron contains truncated forms of both glycoprotein D (gD2) andglycoprotein B (gB2) of HSV-2, purified from transfected Chinese HamsterOvary (CHO) cells and formulated in the adjuvant MF59. Another developedby Glaxo-Smithkline (GSK) contains a truncated gD2 formulated withadjuvants alum and 3-O-deacylated monophosphoryl lipid A (MPL). Bothvaccines were immunogenic and well tolerated in phase I/II trials.However in phase III analyses, the Chiron vaccine showed no overallefficacy against HSV-2 seroconversion and work was discontinued. The GSKvaccine showed significant efficacy (73-74%) in HSV-1, HSV-2seranegative women volunteers but no efficacy in men.

While even limited vaccine efficacy would beneficially impact HSVsufferers, these trials are testing only a small number of vaccinepossibilities. This is because the vaccine discovery has not beensystematic. Pursuance of a whole-virus vaccine assumes that presentationof the pathogen itself to the immune system will generate optimalimmunity. Indeed the breadth and duration of immune responses to wholepathogen vaccines historically have been better than subunit vaccines.However, pathogenicity of the vaccine strain must be considered. Subunitvaccines, to date, have been selected for vaccine testing based on theirassumed importance in disease pathogenesis and immunogenicity duringinfection. These approaches have identified one candidate against HSVwith limited efficacy in some but no efficacy in other formulations.Thus, new and improved methodologies for herpesvirus vaccine discoveryare needed to protect against herpes diseases.

II. SUMMARY OF THE INVENTION

Infection and transmission of HSV-2 is a major health concern. Thepresent disclosure provides, inter alia, certain highly effectivevaccines against HSV-2. Such vaccines can be used either therapeuticallyor prophylactically. The present disclosure also provides specificantigens and methods for using the antigens to elicit an immune responseagainst HSV-2.

In one aspect, the present disclosure describes a vaccine formulationcomprising a pharmaceutical-acceptable carrier and at least onepolypeptide consisting of SEQ ID NOS: 2, 3, 4 and 5 or an immunogenicfragment thereof, and optionally further comprising SEQ ID NO:1 or animmunogenic fragment thereof. The vaccine formulation may comprise afirst polypeptide consisting of one of the above SEQ ID NOS, and asecond polypeptide consisting of another one of the above SEQ ID NOS.

Another aspect of the present invention provides a vaccine formulationcomprising a pharmaceutically acceptable carrier, an adjuvant comprisingone or more purified fractions of quillaja saponins, and at least onepolypeptide comprising any of SEQ ID NOS: 2, 3, 4 and 5 or animmunogenic fragment thereof, and optionally further comprising SEQ IDNO:1 or an immunogenic fragment thereof.

A further aspect of the present invention provides a vaccine formulationcomprising a pharmaceutically-acceptable carrier and a polypeptideconsisting of SEQ ID NO: 2 or an immunogenic fragment thereof. Residuesmay be truncated from SEQ ID NO:2. The polypeptide may be glycosylated,or may be unglycosylated.

In still a further aspect, the present invention provides a vaccineformulation comprising a pharmaceutically-acceptable carrier and apolypeptide comprising SEQ ID NO:5, wherein the polypeptide lacks all orat least an 8 contiguous amino acid residue portion of the transmembranedomain spanning residues 340-363. Accordingly, one aspect of the presentinvention provides a vaccine formulation comprising apharmaceutically-acceptable carrier and a polypeptide comprising SEQ IDNO:4. The polypeptide may be glycosylated, or may be unglycosylated.

Still another aspect of the present invention provides a vaccineformulation comprising a pharmaceutically-acceptable carrier, apolypeptide comprising SEQ ID NO:5. The polypeptide may be glycosylated,or may be unglycosylated.

Yet another aspect of the present invention provides a vaccineformulation comprising a pharmaceutically-acceptable carrier, apolypeptide comprising SEQ ID NO:3. The polypeptide may be glycosylated,or may be unglycosylated.

In some embodiments, polypeptides in the vaccine formulations that maybe conjugated to an immunogenic carrier, for example keyhole limpethemocyanin. In other embodiments, the vaccine formulations furthercomprise an adjuvant. The adjuvant may be one or more purified fractionsof quillaja saponins.

The invention provides methods or treating a subject suffering from orsusceptible to HSV-2 infection by administering an effective amount of avaccine formulation disclosed herein. In some embodiments, the methodinhibits HSV-2 symptoms, for example by reducing the number of herpeticlesions, reducing the number of days a subject experiences herpeticlesions, reducing infection by HSV-2 in an uninfected subject,increasing the IgG titer and/or T cell response to one or more HSV-2antigens, and/or reducing the number of herpetic lesions at the onset ofHSV-2 infection.

In another aspect, the present disclosure describes the results of ahigh-throughput system for in vitro screening of libraries ofefficacious T cells to identify their specific target antigens from thecomplete proteome of HSV-2. This technology allowed the identificationof individual antigens, likely to be effective in vivo, as either aprophylactic or therapeutic composition. In one aspect, herein areprovided several critical protective T cell antigens that can beincorporated into protein-based compositions that elicit an immuneresponse.

One aspect of the present invention provides pharmaceutical compositionscomprising two or more isolated polypeptides selected from polypeptideshaving an amino acid sequence of at least one of SEQ ID NOS: 1-38, or animmunogenic fragment thereof.

In another aspect, the invention provides vaccine formulations thatinclude a pharmaceutically-acceptable carrier and a polypeptidecomprising at least one of SEQ ID NOS: 1-38, or an immunogenic fragmentthereof. In certain embodiments, the polypeptide consists of at leastone of SEQ ID NOS: 1-38.

Another aspect of the present invention provides a method of inducing animmune response in a subject, comprising administering to said subjectan effective amount of a vaccine formulation or a pharmaceuticalcomposition comprising an effective amount of two or more isolatedpolypeptides selected from polypeptides having an amino acid sequence ofat least one of SEQ ID NOS: 1-38, or an immunogenic fragment thereof.

Yet another aspect of the present invention provides a method ofreducing one or more symptoms of HSV-2 infection in a subject,comprising administering to said subject an effective amount of avaccine formulation or a pharmaceutical composition comprising two ormore isolated polypeptides selected from polypeptides having an aminoacid sequence of at least one of SEQ ID NOS: 1-38, or an immunogenicfragment thereof. In some embodiments, the symptoms of HSV-2 infectioncomprise one or more of lesion formation, pain, irritation, itching,fever, malaise, headache, viral shedding, and prodrome.

A further aspect of the present invention provides a method ofinhibiting the onset of HSV-2 infection, comprising administering aneffective amount of a vaccine formulation or a composition comprisingtwo or more isolated HSV polypeptides selected from polypeptides havingan amino acid sequence of at least one of SEQ ID NOS: 1-38, or animmunogenic fragment thereof.

Applicants disclose another aspect of the present invention, whichprovides a method of inhibiting development of a latent HSV-2 infectionin a subject exposed to HSV-2, comprising administering an effectiveamount of a vaccine formulation or a composition comprising two or moreisolated HSV-2 polypeptides selected from polypeptides having an aminoacid sequence of at least one of SEQ ID NOS: 1-38, or an immunogenicfragment thereof.

In a related aspect, the present invention provides a method of reducingviral shedding in a subject infected with HSV-2, comprisingadministering an effective amount of a vaccine formulation or acomposition comprising two or more isolated HSV-2 polypeptides selectedfrom polypeptides having an amino acid sequence of at least one of SEQID NOS: 1-38, or an immunogenic fragment thereof.

Further, an aspect of the present invention provides a method ofreducing recurrence of outbreaks in a subject infected with HSV-2,comprising administering an effective amount of a vaccine formulation ora composition comprising two or more isolated HSV-2 polypeptidesselected from polypeptides having an amino acid sequence of at least oneof SEQ ID NOS: 1-38, or an immunogenic fragment thereof.

An additional aspect of the present invention provides a method ofproducing any of the pharmaceutical compositions described above,comprising expressing said two or more polypeptides; and isolating saidtwo or more polypeptides.

Applicants further disclose an aspect of the present invention whichprovides a method for diagnosing severity of symptoms in a subjectedinfected with HSV-2, comprising (i) measuring activation of T cells inresponse to autologous antigen presenting cells (APC) pulsed with one ormore isolated HSV-2 polypeptides selected from polypeptides set forth inSEQ ID NOS: 1-38, or an immunogenic fragment thereof, and (ii) comparingsaid levels to reference levels obtained from infected subjectsexperiencing frequent outbreaks; whereby a significant increase in saidresponses relative to reference levels indicates that said subject hasless severe symptoms (e.g., the subject is asymptomatic). A significantincrease in response can, for example, comprise a 1.5-fold or greater,2-fold or greater, 3-fold or greater, 5-fold or greater, 10-fold orgreater or even 20-fold or greater increase.

Another aspect of the present invention provides a method for diagnosingseverity of symptoms in a subjected infected with HSV-2, comprising (i)measuring activation of T cells from naturally infected or virus-exposedsubjects in response to APC presenting one or more isolated HSV-2polypeptides selected from polypeptides set forth in SEQ ID NOS: 1-38,or an immunogenic fragment thereof, and (ii) comparing said levels toreference levels obtained from infected subjects experiencing frequentoutbreaks; whereby a significant decrease in said activation relative toreference levels indicates that said subject has more severe symptoms(e.g., frequent outbreaks).

Another aspect of the present invention provides pharmaceuticalcompositions comprising an antibody that binds to one or more isolatedHSV polypeptides selected from the list consisting of SEQ ID NOS: 1-38,or an immunogenic fragment thereof.

Moreover, a different aspect of the present invention provides a methodof identifying immunogenic compositions for HSV-2 by testing two or morepolypeptides selected from polypeptides having an amino acid sequence ofany one of SEQ ID NOs. 1-38, or an immunogenic fragment thereof, forability to promote cytokine production in a mammalian T cell, wherein animmunogenic composition is one that elevates levels of a cytokinesignificantly above the levels of that cytokine produced by a naïvemammalian T cell. A significant increase in cytokine levels is typicallyone that is at least 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold or even20-fold the level produced by a naïve cell.

Still another aspect of the present invention provides a method ofdetecting HSV-2 in a sample from a subject, said method comprising (i)contacting said sample with one or more antibodies raised against one ormore polypeptides having an amino acid sequence of SEQ ID NOS: 1-38 oran immunogenic fragment thereof, and (ii) detecting said one or moreantibodies bound to said one or more HSV-2 polypeptide from the sample.

Finally, one aspect of the present invention provides pharmaceuticalcompositions comprising two or more isolated polynucleotides, selectedfrom nucleotide SEQ ID NOS: 1-38, or fragments encoding immunogenicpeptides thereof.

III. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A and FIG. 1B are graphs showing, respectively, CD4⁺ and CD8⁺ Tcell responses following immunization with gD2 full-length protein,gD2ΔTMR, or gD2 truncated immediately upstream of the transmembranedomain (denoted 306t).

FIG. 2A and FIG. 2B are graphs showing, respectively, CD4⁺ and CD8⁺ Tcell responses following immunization with pooled, overlapping peptidesspanning gL2, or ICP4 fragments encoded by RS1.1, RS1.3.1 and RS1.3.2.

FIG. 3A and FIG. 3B are graphs showing, respectively, CD4⁺ and CD8⁺ Tcell responses following immunization with gD2ΔTMR, or gD2ΔTMR andICP4.2.

IV. DETAILED DESCRIPTION

This application describes vaccines and immunogenic compositions againstHSV-2. Vaccine formulations may include a polypeptide comprising asequence from Table 1 or an immunogenic fragment thereof, or acombination of at least two polypeptides comprising sequences from Table1 or immunogenic fragments thereof. In certain embodiments, thepolypeptide(s) of the vaccines comprise the entire sequence of at leastone of SEQ ID NOS: 1-26 or consist of the entire sequence of any one ofSEQ ID NOS: 1-26. Immunogenic compositions may include a polypeptidecomprising a sequence from Table 1 or Table 2 or an immunogenic fragmentthereof or a combination of at least two polypeptides comprisingsequences from Table 1 or Table 2, or immunogenic fragments thereof. Incertain embodiments, the polypeptide(s) of the immunogenic compositionscomprise the entire sequence of any one of SEQ ID NOS: 1-38 or consistof the entire sequence of SEQ ID NO: 1-38. The polypeptides in Tables 1or 2 may be encoded by SEQ ID NOS: 39-46 and 117-134 as indicated and/orby cDNA sequences publically available onhttp://www.ncbi.nlm.nih.gov/sites/entrez. cDNA and protein sequences mayalso be obtained from any known strains of HSV-2, including HG52, 333,and Strain G. Accordingly, cDNA sequences may be accessed by gene orprotein name from genomic sequence at NC_001798.1, and may beapproximately 97% conserved with sequences disclosed at NC_001798.1). Asdescribed herein, the polypeptides may be referred to by protein name,by SEQ ID NO, and/or by the name of the gene encoding the protein.

The polypeptides can be prepared in a variety of expression systems.Suitable expression systems include E. coli and Baculovirus-basedexpression systems (e.g., in insect cells). Polypeptides prepared usingE. coli are typically full-length and unglycosylated, although truncatedvariants can be prepared. In certain embodiments, these truncatedvariants retain all or part of the signal domain. Polypeptides preparedusing a Baculovirus system typically lack the N-terminal signalsequence, but are fully or partially glycosylated.

TABLE 1 HSV-2 antigens for vaccines or immunogenic compositions ProteinDNA SEQ ID SEQ ID Gene Name Gene GenBank No. No. Protein Name ID No.Accession Nos. 1 39 RS1 1869897 NP_044530.1 ICP4 2 117 RS1.2 RS1.2corresponds to an ICP4 internal internal fragment of the RS1 fragment(ICP4.2) sequence 3 118 UL1 1487292 NP_044470.1 gL2 cytoplasmic 4 40US6ΔTMR 9629336 NP_044536.1 gD2 internal US6ΔTMR corresponds to deletion(gDΔTMR) gD2 with a deletion of amino acids 340-363 5 US6 gD2 6 41 RL19629329 NP_044529.1 ICP34.5 7 42 RL2 109676722 NP_044528.2 ICP0 8 121RS1.1 1869897 NP_044530.1 ICP4 internal RS1.1 corresponds to residuesfragments 1-400 of RS1 9 122 RS1.3.1 1869897 NP_044530.1 ICP4 internalRS 1.3.1 corresponds to fragments residues 750-1024 of RS1 10 123RS1.3.2 1869897 NP_044530.1 ICP4 internal RS1.3.2 corresponds tofragments residues 1008-1319 of RS1 11 124 RS1.3 1869897 NP_044530.1ICP4 internal RS1.3 corresponds to residues fragments 750-1319_of RS1 12125 RS1.4 1869897 NP_044530.1 ICP4 internal RS1.4 corresponds toresidues fragments 340-883 of RS1 13 126 RS1.5 1869897 NP_044530.1 ICP4internal RS1.5 corresponds to residues fragments 775-1318 of RS1 14 127RS1.6 1869897 NP_044530.1 ICP4 internal RS1.6 corresponds to residuesfragments 209-1318 of RS1 15 128 RS1.7 1869897 NP_044530.1 ICP4 internalRS1.7 has a deletion of fragments residues 391-544 of RS1 16 129 RS1.81869897 NP_044530.1 ICP4 internal RS1.8 has a deletion of fragmentsresidues 786-864 of RS1 17 UL2 uracil DNA glycosylase 18 UL11myristylated tegument protein 19 119 UL1 1487292 NP_044470.1 gL2secreted 20 UL19 VP5 21 120 UL19ΔTEV 9629288 NP_044488.1 VP5 UL19ΔTEV islacking the last 5 amino acids from the C- terminal end of UL19 22 UL36ICP½ 23 43 UL36.3.4.1 1487322 NP_044506.1 ICP½ internal UL 36.3.4.1corresponds to fragments residues 1318-2280 of UL36 24 44 UL36.4.2.51487322 NP_044506.1 ICP½ internal UL 36.4.2.5 corresponds to fragmentsresidues 2253-3122 of UL36 25 UL40 ribonucleoside reductase 26 45 US129629343 NP_044543.1 ICP47

TABLE 2 Additional HSV-2 antigens for immunogenic compositions ProteinDNA SEQ ID SEQ ID Gene Name Gene GenBank No. No. Protein Name ID No.Accession Nos. 27 134 UL10 9629279 NP_044479.1 gM2 28 UL15 DNAcleavage/packaging protein 29 UL26.5 ICP35 30 UL30 DNA-directedpolymerase 31 UL5 DNA helicase/primase complex 32 UL8 DNAhelicase/primase complex 33 UL15.5 unknown 34 UL32 cleavage andpackaging protein 35 UL36.4.2 ICP½ fragment 36 UL54 ICP27 37 133 UL49.51487337 NP_044520.1 Membrane associated virion protein 38 46 US4 9629334NP_044534.1 gG2A. Immunogenic HSV-2 Polypeptides

Immunogenic polypeptides or polynucleotides as indicated in Table 1and/or Table 2 may be used in pharmaceutical compositions. The inventionprovides pharmaceutical compositions containing immunogenic polypeptidesor polynucleotides encoding these immunogenic polypeptides together witha pharmaceutical carrier. Antigens from HSV-2 may be identified byscreening immune cells from patients infected with HSV-2. Briefly, alibrary of HSV-2 antigens was expressed by bacteria and mixed withantigen presenting cells (APCs). The APCs, in turn, processed andpresented HSV-2-derived polypeptides to lymphocytes that had beenisolated from human patients infected with HSV-2. The patients belongedto several populations: (1) exposed to HSV-2 but seronegative forinfection, (2) infected with HSV-2 but asymptomatic, (3) infected withHSV-2 and experiencing infrequent outbreaks, (4) infected with HSV-2 andexperiencing frequent outbreaks, (5) naïve and (6) seronegative forHSV-2 (HSV-2⁻) but seropositive for HSV-1 (HSV-1⁺). Lymphocyte responsesfrom each population were compared for reactivity to HSV-2-derivedpolypeptides, and the screen detected antigens that induced reactivelymphocytes with greater frequency in one patient population as comparedto the others. Infected but asymptomatic, and exposed but seronegativepatients may activate protective immune responses that patients whoexperience frequent outbreaks do not; in particular, exposed butseronegative patients are presumed to have mounted sterilizing immunityto HSV-2 infection. It is believed that a unique set of polypeptideswill activate lymphocytes from these patient populations. Thus, thepresent invention contemplates compositions of the specific HSV-2polypeptides that activate the lymphocytes of infected but asymptomatic,or exposed but seronegative patients or a combination of thesepolypeptides for inhibiting or counteracting infection by HSV-2.

Antigens identified on the basis of their immunogenicity in infected butasymptomatic, or exposed but seronegative patients are similarlyexpected to be immunogenic in any subject.

In some embodiments, a polypeptide may induce an innate immune response,a humoral immune response, or a cell-mediated immune response. Thecell-mediated immune response may involve T_(H)1 cells, and in certainembodiments, the immune response involving T_(H)1 cells is an immuneresponse in which T_(H)1 cells are activated. In some embodiments, animmunogenic polypeptide avoids induction of T_(H)2 cytokines. In someembodiments, the cell-mediated immune response may involve T_(H)17cells, and in certain embodiments, the immune response involving T_(H)17cells is an immune response in which T_(H)17 cells are activated.

Polypeptides (or immunogenic fragments thereof) in compositions of theinvention may induce T cell responses in multiple individuals,regardless of the HLA haplotype of the individuals. Specifically,epitopes on the polypeptides may induce T cell responses in individualswith one or more of the following HLA supertypes: HLA-A2, -A3, -A24,-A1, -B7, -B8, -B27, -B44, -B58, and B62, and HLA-DQB01, -DQB02, -DQB03,-DQB-04, and -DQB05.

In some embodiments, one or more, e.g. two, three, four, or morepolypeptides from Table 1 and/or Table 2 (or immunogenic fragmentsthereof) are provided in a composition of the invention. In someembodiments, two polypeptides from Table 1 and/or Table 2 are providedin a composition of the invention. In other embodiments, threepolypeptides from Table 1 and/or Table 2 are provided in a compositionof the invention.

In some embodiments, two, three, four, or more polypeptides from Table 1and/or Table 2 (or immunogenic fragments thereof) are provided togetheras a conjugate. In some embodiments, two polypeptides from Table 1and/or Table 2, or three polypeptides from Table 1 and/or Table 2, areprovided as a conjugate. In some embodiments, two, three, four, or morepolypeptides from Table 1 and/or Table 2 are covalently bound to eachother, e.g., as a fusion protein. In some embodiments, two, three, four,or more polypeptides from Table 1 and/or Table 2 are covalently bound toeach other, e.g., as a fusion protein. In some embodiments, twopolypeptides from Table 1 and/or Table 2, or three polypeptides fromTable 1 and/or Table 2, are covalently bound to each other, e.g. as afusion protein.

In some embodiments, the compositions comprise two or more polypeptidesselected from the group consisting of SEQ ID Nos. 1-38, and may containor may not contain any other HSV-2 polypeptides.

In certain embodiments, Applicants provide polypeptides that are atleast 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to apolypeptide encoded by a gene in Table 1 and/or Table 2, or a portion ofsaid polypeptide. In certain embodiments, the homologous polypeptide isat least 8, 10, 15, 20, 30, 40, 50, 60, 80, 100, 120, 140, 160, 180,200, 220, 240, 260, 280, 300, 350, 400, 450, or 500 amino acids inlength. In some embodiments, such as those described immediately above,the polypeptide is no more than 300, 350, 400, 450, or 500 amino acidsin length.

An immunogenic composition may also comprise portions of saidpolypeptides and genes, for example deletion mutants, truncationmutants, oligonucleotides, and peptide fragments. In some embodiments,the portions of said proteins are immunogenic.

The immunogenicity of a portion of a protein or a homolog thereof can bereadily determined using the same assays that are used to determine theimmunogenicity of the full-length protein. In some embodiments, theportion of the protein has substantially the same immunogenicity as thefull-length proteins. In some embodiments, the immunogenicity is no morethan 10%, 20%, 30%, 40%, or 50% less than that of the full-lengthprotein. The protein fragments may be, for example, linear, circular, orbranched. In some embodiments, a protein or protein fragment comprisesone or more non-natural amino acids (e.g. an amino acid other than the20 typically found in natural proteins). A non-natural amino acid mayhave an atypical side chain. In addition, peptidomimetics may be used;these may incorporate alterations to the peptide backbone.

Some embodiments of the polypeptide composition described herein includean immunogenic polypeptide that contains a membrane translocatingsequence (MTS), to facilitate introduction of the polypeptide into themammalian cell and subsequent stimulation of the cell-mediated immuneresponse. Exemplary membrane translocating sequences include hydrophobicregion in the signal sequence of Kaposi fibroblast growth factor, theMTS of α-synuclein, β-synuclein, or γ-synuclein, the third helix of theAntennapedia homeodomain, SN50, integrin β3 h-region, HIV Tat, pAntp,PR-39, abaecin, apidaecin, Bac5, Bac7, P. berghei CS protein, and thoseMTSs described in U.S. Pat. Nos. 6,248,558, 6,432,680 and 6,248,558.

In certain embodiments, the immunogenic polypeptide is conjugated (i.e.covalently bound) to another molecule. This may, for example, increasethe half-life, solubility, bioavailability, or immunogenicity of theantigen. Molecules that may be conjugated to an immunogenic polypeptideinclude a carbohydrate, biotin, poly(ethylene glycol) (PEG), polysialicacid, N-propionylated polysialic acid, nucleic acids, polysaccharides,and PLGA. There are many different types of PEG, ranging from molecularweights of below 300 g/mol to over 10,000,000 g/mol. PEG chains can belinear, branched, or with comb or star geometries.

B. Immunogenic HSV-2 Polypeptides and Nucleic Acids for Use in Vaccines

In certain embodiments, one or more, e.g. two, three, four, or moreimmunogenic fragments or variants thereof are provided in a mixture. Forexample, a vaccine formulation may comprise any one or more of SEQ IDNOS: 1-26.

In certain embodiments, a vaccine formulation may comprise any one, two,or three of ICP4, ICP4.2, gL2, gD2ΔTMR and gD2 (SEQ ID NOS: 1-5), orimmunogenic fragment(s) thereof. In certain embodiments, combinationscontain polypeptides or immunogenic fragments from only one of ICP4 (SEQID NO 1) and ICP4.2 (SEQ ID NO 2). In other embodiments, combinationscontain polypeptides or immunogenic fragments from only one of gD2ΔTMR(SEQ ID NO:4) and gD2 (SEQ ID NO:5).

Exemplary combinations of ICP4, ICP4.2, gL2, gD2ΔTMR and gD2 include:

Two antigen combinations ICP4 gL2 SEQ ID NO: 1 SEQ ID NO: 3 ICP4 gD2ΔTMRSEQ ID NO: 1 SEQ ID NO: 4 ICP4 gD2 SEQ ID NO: 1 SEQ ID NO: 5 ICP4.2 gL2SEQ ID NO: 2 SEQ ID NO: 3 ICP4.2 gD2ΔTMR SEQ ID NO: 2 SEQ ID NO: 4ICP4.2 gD2 SEQ ID NO: 2 SEQ ID NO: 5 gL2 gD2ΔTMR SEQ ID NO: 3 SEQ ID NO:4 gL2 gD2 SEQ ID NO: 3 SEQ ID NO: 5

Three antigen combinations ICP4 gL2 gD2ΔTMR SEQ ID NO: 1 SEQ ID NO: 3SEQ ID NO: 4 ICP4.2 gL2 gD2ΔTMR SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4ICP4 gL2 gD2 SEQ ID NO: 1 SEQ ID NO: 3 SEQ ID NO: 5 ICP4.2 gL2 gD2 SEQID NO: 2 SEQ ID NO: 3 SEQ ID NO: 5

The individual antigens and combinations described above can alsoinclude additional peptides from or derived from HSV-2, such aspolypeptides comprising sequences selected from SEQ ID NO:6-26 orimmunogenic fragments thereof.

1. ICP4 (SEQ ID NO: 1) Encoded by RS1

RS1 encodes ICP4, a transcriptional transactivator that may interactwith and recruit specific components of the general transcriptionmachinery to viral promoters and stabilize their formation fortranscription initiation. ICP4 contains distinct domains fortransactivation/phosphorylation (approximately spanning acid residues150-200 of SEQ ID NO:1), DNA binding (approximately spanning residues380-540 of SEQ ID NO:1), nuclear localization (approximately spanningresidues 630-730 of SEQ ID NO:1), and late regulatory transactivation(approximately spanning residues 1220-1319 of SEQ ID NO:1). The DNA andprotein sequence of RS1 may be found by searching for RS1 in thepublicly available database, Entrez Gene (on the NCBI NIH web site onthe World Wide Web, at www.ncbi.nlm.nih.gov/sites/entrez?db=gene), inthe Human herpesvirus 2 complete genome.

no more than 1000 amino acids of ICP4 (SEQ ID NO: 1). The polypeptidemay also be a variant of the at least 20 residue fragment.

In certain embodiments, the polypeptide includes no more than 950, 900,850, 800, 750, 700, 650, 600, 550, 500, 450 or even 400 consecutiveamino acids from ICP4. Exemplary polypeptides correspond approximatelyto amino acids residues of full-length ICP4 as follows: 383-766; 1-400(RS1.1); 750-1024 (RS1.3.1); 1008-1319 (RS1.3.2); 750-1319 (RS1.3);280-785 (RS1.4 comprising the full DNA binding region); 680-1319 (RS1.5comprising the glycosylase/C-terminal region); 208-1319 (RS1.6 which mayalso comprise a Met residue at the N-term end); 1-380 plus 545-1319(RS1.7, in which a region spanning approximately residues 381-544 isdeleted, removing the DNA binding regions); 1-785 plus 870-1319 (RS1.8,in which a region spanning approximately residues 786-869 is deleted,removing the nuclear localization domain), or 1-766, 383-1318, 100-750,400-1300, 250-766, 383-900 of ICP4 (SEQ ID NO. 1) and the like.

2. ICP4 Internal Fragment ICP4.2 (SEQ ID NO: 2) Encoded by RS1.2

RS1.2 encodes a 391 amino acid fragment of ICP4 denoted ICP4.2.

In specific embodiments, vaccines against HSV-2 include a polypeptidecontaining from 50 to all 391 amino acids residues of ICP4.2 (SEQ ID NO:2), such as from 100 to 391, 200 to 391 or 250 to 350 residues. Inparticular embodiments, the polypeptide includes all of ICP4.2 (SEQ IDNO: 2) or is ICP4.2 (SEQ ID NO: 2) itself. These polypeptides may, forexample, include the full length or fragments of ICP4.2 (SEQ ID NO:2)described herein with amino acids residues 1-382 or 767-1318 of ICP4(SEQ ID NO. 1) or fragments thereof, which, in certain embodiments, areconsecutive with the amino acid residues of ICP4.2 being used. Exemplaryfragments that combine the residues of SEQ ID NO:2 with select residuesfrom 1-382 or 767-1318 of SEQ ID NO:1 are described above.

An immunogenic fragment of ICP4.2 comprises at least one immunogenicportion, as measured experimentally or identified by algorithm. Peptidesidentified by such methods include the following:

(SEQ ID NO: 47) GLAHVAAAV (SEQ ID NO: 48) FISGSVARA (SEQ ID NO: 49)QYALITRLL (SEQ ID NO: 50) RYDRAQKGF (SEQ ID NO: 51) GYAMAAGRF(SEQ ID NO: 52) PPHADAPRL (SEQ ID NO: 53) KPAAAAAPL (SEQ ID NO: 54)SEAAVAAV (SEQ ID NO: 55) FGWGLAHV (SEQ ID NO: 56) YALITRLLY(SEQ ID NO: 57) ALPRSPRLL (SEQ ID NO: 58) DLLFQNQSL (SEQ ID NO: 59)ADLLFQNQS (SEQ ID NO: 60) ARNSSSFIS (SEQ ID NO: 61) QACFRISGA(SEQ ID NO: 62) FVRDALVLM (SEQ ID NO: 63) FDGDLAAVP (SEQ ID NO: 64)GLGDSRPGL (SEQ ID NO: 65) WAPELGDAA (SEQ ID NO: 66) ECLAACRGI(SEQ ID NO: 67) RAWLRELRF.

Thus, in some aspects, this application provides an immunogenic fragmentof ICP4.2. The fragments, in some instances, are close in size to thefull-length polypeptide. For example, they may lack at most one, two,three, four, five, ten, or twenty amino acids from one or both termini.In other embodiments, the fragment is 100-391 amino acids in length, or150-391, or 200-391, or 250-391 amino acids in length. Other exemplaryfragments are amino acid residues 1-350, 1-300, 1-250, 1-200, 1-150,1-100, 1-50, 50-391, 50-350, 50-300, 50-250, 50-200, 50-150, 50-100,100-391, 100-350, 100-300, 100-250, 100-200, 100-150, 150-391, 150-350,150-300, 150-250, 150-200, 200-391, 200-350, 200-300, 200-250, 250-391,250-350, 250-300, 300-391 and 350-391. The fragments described above orsub-fragments thereof (e.g., fragments of 8-50, 8-30, or 8-20 amino acidresidues) preferably have one of the biological activities describedbelow, such as increasing the T cell response by at least 1.5 fold or 2fold. A fragment may be used as the polypeptide in the vaccinesdescribed herein or may be fused to another protein, protein fragment ora polypeptide.

In certain aspects, this application provides immunogenic polypeptideswith at least 90%, 95%, 97%, 98%, 99%, or 99.5% identity to ICP4.2 or animmunogenic fragment thereof.

3. Glycoprotein L-2 (SEQ ID NO: 3) Encoded by UL1

UL1 encodes Glycoprotein L-2 (gL2), a heterodimer glycoprotein that isrequired for the fusion of viral and cellular membranes and enables thevirus to enter the host cell. The DNA and protein sequence of UL1 may befound by searching in the publicly available database, Entrez Gene (onthe NCBI NIH web site on the World Wide Web, atww.ncbi.nlm.nih.gov/sites/entrez?db=gene), in the Human herpesvirus 2complete genome.

In some embodiments, vaccines against HSV-2 include a polypeptidecontaining at least 20 consecutive amino acid residues selected fromresidues 1-224 of gL2 (SEQ ID NO: 3), but no more than 224 amino acidsof gL2 (SEQ ID NO: 3). The polypeptide may also be a variant of the atleast 20 residue fragment.

In some embodiments, the polypeptide is at least 85% identical to afragment of 200-250 amino acids of SEQ ID NO: 3.

In certain embodiments, the polypeptide includes no more than 200 or 100consecutive amino acids from gL2. Exemplary polypeptides are amino acidsresidues 1-20, 21-40, 41-60, of 61-80, 81-100, 101-120, 121-140,141-160, 161-180, 181-200, 201-221 of gL2 (SEQ ID NO. 3) and the like.

In other aspects, this application provides an immunogenic fragment ofgL2. An immunogenic fragment of gL2 comprises at least one immunogenicportion, as measured experimentally or identified by algorithm. Peptidesidentified by such methods include the following:

(SEQ ID NO: 100) AYLVNPFLF (SEQ ID NO: 101) PFLFAAGFL (SEQ ID NO: 102)TEYVLRSVI (SEQ ID NO: 103) GSQATEYVL (SEQ ID NO: 104) RIDGIFLRY(SEQ ID NO: 105) FLEDLSHSV (SEQ ID NO: 106) YVLRSVIAK (SEQ ID NO: 107)YVLRSVIAK (SEQ ID NO: 108) AYLVNPFLF (SEQ ID NO: 109) ETTTRRALY(SEQ ID NO: 110) RIDGIFLRY (SEQ ID NO: 111) YLVNPFLFA (SEQ ID NO: 112)FVCLFGLVV (SEQ ID NO: 113) LYKEIRDAL (SEQ ID NO: 114) GLDTFLWDR(SEQ ID NO: 115) RVSPTRGRR (SEQ ID NO: 115) YVLRSVIAK (SEQ ID NO: 116)GLDTFLWDR (SEQ ID NO: 117) DILRVPCMR (SEQ ID NO: 118) DRHAQRAYL4. Glycoprotein D-2 (SEQ ID NO: 5) Encoded by US6 and Internally-DeletedGlycoprotein D-2 (SEQ ID NO:4) Encoded by US6ΔTMR

US6 encodes envelope glycoprotein D-2 (gD2), an envelope glycoproteinthat binds to host cell entry receptors and may trigger fusion of thevirus with the host membrane. The gD2 protein has several distinctdomains, including a signal domain (amino acid residues 1-25) which iscleaved from the mature protein, and a transmembrane domain (spanningapproximately amino acids residues 340-363). The DNA and proteinsequence of US6 may be found by searching in the publicly availabledatabase, Entrez Gene (on the NCBI NIH web site on the World Wide Web,at www.ncbi.nlm.nih.gov/sites/entrez?db=gene), in the Human herpesvirus2 complete genome.

In some embodiments, vaccines against HSV-2 include a polypeptidecomprising gD2 that is missing all or part of the transmembrane domain(which spans approximately amino acids residues 340-363 inclusive) aswell as the signal sequence. In other embodiments, the deleted regionmay additionally include 5-10 amino acids of the sequence flanking thetransmembrane domain. The deleted region may also comprise a portion ofthe transmembrane domain, for example at least 3 amino acids betweenresidues 340-363. In some embodiments, at least one residue in thetransmembrane domain has been modified, deleted or substituted, suchthat the transmembrane domain is no longer functional. For example, avariant may have its internal deletion begin at amino acid residue 336,337, 338, 339, 340, 341, 342, 343, 344, 345 or 346 and end at amino acidresidue 358, 359, 360, 361, 362, 363, 364, 365, 366, 367 or 368.

A construct encoding gD2 which is missing amino acid residues 340-363(the transmembrane domain) is called US6ΔTMR (SEQ ID NO: 40). Thecorresponding protein is denoted gD2ΔTMR (SEQ ID NO:4). In otherembodiments, an immunogenic fragment of gD2 or gD2ΔTMR may comprise adeletion in a portion of the transmembrane domain, and/or may comprise adeletion in the flanking sequence outside of the transmembrane domain.

In other aspects, this application provides an immunogenic fragment ofgD2 or gD2ΔTMR. An immunogenic fragment of gD2 or gDΔTMR comprises atleast one immunogenic portion, as measured experimentally or identifiedby algorithm. Peptides identified by such methods include the following:

(SEQ ID NO. 68) ALAGSTLAV (SEQ ID NO. 69) LLEDPAGTV (SEQ ID NO. 70)VIGGIAFWV (SEQ ID NO. 71) TVYYAVLER (SEQ ID NO. 72) KYALADPSL(SEQ ID NO. 73) AFETAGTYL (SEQ ID NO. 74) APSNPGLII (SEQ ID NO. 75)IPITVYYAV (SEQ ID NO. 76) APPSHQPLF (SEQ ID NO. 77) FLMHAPAFE(SEQ ID NO. 78) FSAVSEDNL (SEQ ID NO. 79) VYYAVLER (SEQ ID NO. 80)IGMLPRFI (SEQ ID NO. 81) YTECPYNKS (SEQ ID NO. 82) FLMHAPAFE(SEQ ID NO. 83) NLGFLMHAP (SEQ ID NO. 84) VIGGIAFWV (SEQ ID NO. 85)GIAFWVRRR (SEQ ID NO. 86) SEDNLGFLM (SEQ ID NO. 87) RTQPRWSYY(SEQ ID NO. 88) IAFWVRRRA (SEQ ID NO. 89) LVIGGIAFW (SEQ ID NO. 90)FWVRRRAQM (SEQ ID NO. 91) PYTSTLLPP (SEQ ID NO. 92) VGTAALLVV(SEQ ID NO. 93) TAALLVVAV (SEQ ID NO. 94) TSTLLPPEL (SEQ ID NO. 95)GTVSSQIPP (SEQ ID NO. 96) TAGTYLRLV (SEQ ID NO. 97) GVTVDSIGM(SEQ ID NO. 98) AFWVRRRAQ (SEQ ID NO. 99) RVYHIQPSL

Thus, in some aspects, this application provides an immunogenic fragmentof gD2 (SEQ ID NO:5) or gDΔTMR (SEQ ID NO: 4). The fragments, in someinstances, are close in size to the full-length polypeptide. Forexample, they may lack at most one, two, three, four, five, ten, ortwenty amino acids from one or both termini. In other embodiments, thefragment is 100-393 amino acids in length, or 150-393, or 200-393, or250-393 amino acids in length. Other exemplary fragments are amino acidresidues 1-350, 1-300, 1-250, 1-200, 1-150, 1-100, 1-50, 50-393, 50-350,50-300, 50-250, 50-200, 50-150, 50-100, 100-393, 100-350, 100-300,100-250, 100-200, 100-150, 150-393, 150-350, 150-300, 150-250, 150-200,200-393, 200-350, 200-300, 200-250, 250-393, 250-350, 250-300, 300-393and 350-393. The fragments described above or sub-fragments thereof(e.g., fragments of 8-50, 8-30, or 8-20 amino acid residues) preferablyhave one of the biological activities described below, such asincreasing the T cell response by at least 1.5 fold or 2 fold. Afragment may be used as the polypeptide in the vaccines described hereinor may be fused to another protein, protein fragment or a polypeptide.

In other embodiments, the polypeptide comprises the entire sequence ofSEQ ID NO: 4 or SEQ ID NO:5, or consists of the entire sequence of SEQID NO: 4 or SEQ ID NO:5. In certain embodiments, an immunogenic fragmentof gD2 retains all or part of the signal domain (amino acid residues1-25) and/or the transmembrane domain (amino acids residues 340-363).

In certain embodiments, polypeptides have less than 20%, 30%, 40%, 50%,60% or 70% homology with human autoantigens. Examples of suchautoantigens include UL6 from HSV-1 and gK or UL53 from HSV-2.

In certain aspects, this application provides immunogenic polypeptideswith at least 90%, 95%, 97%, 98%, 99%, or 99.5% identity to gDΔTMR, oran immunogenic fragment thereof.

C. Additional Features of HSV-2 Polypeptides

Typically, the polypeptides present in the vaccine formulations orpharmaceutical compositions described herein are immunogenic, eitheralone or as a variant, which includes polypeptides fused to anotherpolypeptide or mixed with or complexed to an adjuvant. Variants alsoinclude sequences with less than 100% sequence identity, as describedherein. In addition, one may use fragments, precursors and analogs thathave an appropriate immunogenicity.

These polypeptides may be immunogenic in mammals, for example, mice,guinea pigs, or humans. An immunogenic polypeptide is typically onecapable of raising a significant immune response in an assay or in asubject. Alternatively, an immunogenic polypeptide may (i) induceproduction of antibodies, e.g., neutralizing antibodies, that bind tothe polypeptide (ii) induce T_(H)1 immunity, (iii) activate the CD8+ CTLresponse, for example by increasing CD8+ T cells and/or increasinglocalization of CD8+ T cells to the site of infection or reinfection,(iv) induce T_(H)17 immunity, and/or (v) activate innate immunity. Insome embodiments, an immunogenic polypeptide causes the production of adetectable amount of antibody specific to that antigen.

In certain embodiments, polypeptides have less than 20%, 30%, 40%, 50%,60% or 70% homology with human autoantigens.

A polypeptide may comprise one or more immunogenic portions and one ormore non-immunogenic portions. The immunogenic portions may beidentified by various methods, including protein microarrays,ELISPOT/ELISA techniques, and/or specific assays on different deletionmutants (e.g., fragments) of the polypeptide in question. Immunogenicportions may also be identified by computer algorithms. Some suchalgorithms, like EpiMatrix (produced by EpiVax), use a computationalmatrix approach. Other computational tools for identifying antigenicepitopes include PEPVAC (Promiscuous EPitope-based VACcine, hosted byDana Farber Cancer Institute on the world wide web atimmunax.dfci.harvard.edu/PEPVAC), MHCPred (which uses a partial leastsquares approach and is hosted by The Jenner Institute on the world wideweb at www.jenner.ac.uk/MHCPred), and Syfpeithi, hosted on the worldwide web at www.syfpeithi.de/.

In some embodiments, the vaccine or pharmaceutical composition maycomprise fusion proteins and/or fusion DNA constructs. The underlyingDNA sequences above may be modified in ways that do not affect thesequence of the protein product. For instance, the DNA sequence may becodon-optimized to improve expression in a host such as E. coli or aninsect cell line (e.g. using the baculovirus expression system) ormammalian (e.g. Chinese Hamster Ovary) cell line. In particularembodiments, such as when smaller related polypeptides, including thosehaving a molecular weight less than about 5000 daltons, e.g., 1500 to5000 daltons, are used, modification may be useful in eliciting thedesired immune response. For example, the smaller polypeptides can beconjugated to an appropriate immunogenic carrier such as proteins fromother pathogenic organisms or viruses (e.g., tetanus toxoid), largeproteins (e.g., keyhole limpet hemocyanin) or the like. Conjugation maybe direct or indirect (e.g., via a linker). In other particularembodiments, a fusion protein may comprise a polypeptide disclosed aboveor an immunogenic fragment or variant thereof and a tag. A tag may beN-terminal or C-terminal. For instance, tags may be added to the nucleicacid or polypeptide to facilitate purification, detection, solubility,or confer other desirable characteristics on the protein or nucleicacid. For instance, a purification tag may be a peptide, oligopeptide,or polypeptide that may be used in affinity purification. Examplesinclude His, GST, TAP, FLAG, myc, HA, MBP, VSV-G, thioredoxin, V5,avidin, streptavidin, BCCP, Calmodulin, Nus, S tags, lipoprotein D, andβ galactosidase. In some embodiments, the fused portion is short. Thus,in some instances, the fusion protein comprises no more than 1, 2, 3, 4,5, 10, 20, or 50 additional amino acids on one or both termini of apolypeptide described above, such as consecutive amino acids from any ofthe polypeptides in Table 1.

In some embodiments, tags, secretion signals, or other signal sequencesmay be added to the C-terminal end and/or to the N-terminal end of thepolypeptide. Tags may be used to aid in purification of expressedpolypeptides. Exemplary tags include HHHHHH (SEQ ID NO: 130) andMSYYHHHHHH (SEQ ID NO: 131). Secretion signals may be optimized for usewith non-mammalian cells, such as insect cells. An exemplary secretionsignal is MKFLVNVALVFMVVYISYIYA (SEQ ID NO: 132).

A detection tag may be used to detect the tag and, consequently, anyamino acid sequence fused to it. Detection tags include fluorescentproteins, proteins that bind a fluorescent label, and proteins that bindan electron-dense moeity. Examples of fluorescent proteins includedsRed, mRFP, YFP, GFP, CFP, BFP, and Venus. An example of a protein thatbinds a fluorescent or electron-dense label is FlAsH.

Another aspect disclosed herein is an antibody preparation generatedagainst a composition of the invention (e.g., a composition comprisingone or more or two or more of the polypeptides listed in Table 1). Anyof a variety of antibodies are included. Such antibodies include, e.g.,polyclonal, monoclonal, recombinant, humanized or partially humanized,single chain, Fab, and fragments thereof, etc. The antibodies can be ofany isotype, e.g., IgA, IgG, various IgG isotypes such as IgG₁, IgG₂,IgG_(2a), IgG_(2b), IgG₃, IgG₄, etc.; and they can be from any animalspecies that produces antibodies, including goat, rabbit, mouse, chickenor the like. In some embodiments, Fab molecules are expressed andassembled in a genetically transformed host like E. coli. A lambdavector system is available thus to express a population of Fab's with apotential diversity equal to or exceeding that of subject generating thepredecessor antibody. See Huse et al. (1989), Science 246, 1275-81.

D. Components of Vaccines and Pharmaceutical Compositions

In certain embodiments, the vaccines and pharmaceutical compositionscomprise one or more of the polypeptides and nucleic acids describedabove and one or more of the following: an adjuvant, stabilizer, buffer,surfactant, controlled release component, salt, preservative, and anantibody specific to said antigen.

1. Adjuvants

The vaccine formulations and pharmaceutical compositions describedherein may each include an adjuvant. Adjuvants can be broadly separatedinto two classes, based on their principal mechanisms of action: vaccinedelivery systems and immunostimulatory adjuvants (see, e.g., Singh etal., Curr. HIV Res. 1:309-20, 2003). Vaccine delivery systems are oftenparticulate formulations, e.g., emulsions, microparticles,immune-stimulating complexes (ISCOMs), which may be, for example,particles and/or matrices, and liposomes. In contrast, immunostimulatoryadjuvants are sometimes derived from pathogens and can representpathogen associated molecular patterns (PAMP), e.g., lipopolysaccharides(LPS), monophosphoryl lipid (MPL), or CpG-containing DNA, which activatecells of the innate immune system.

Alternatively, adjuvants may be classified as organic and inorganic.Inorganic adjuvants include alum salts such as aluminum phosphate,amorphous aluminum hydroxyphosphate sulfate, and aluminum hydroxide,which are commonly used in human vaccines. Organic adjuvants compriseorganic molecules including macromolecules. An example of an organicadjuvant is cholera toxin.

Adjuvants may also be classified by the response they induce, andadjuvants can activate more than one type of response. In someembodiments, the adjuvant induces the activation of CD4+ T cells. Theadjuvant may induce activation of T_(H)1 cells and/or activation ofT_(H)17 cells and/or activation of T_(H)2 cells. Alternately, theadjuvant may induce activation of T_(H)1 cells and/or T_(H)17 cells butnot activation of T_(H)2 cells, or vice versa. In some embodiments, theadjuvant induces activation of CD8+ T cells. In further embodiments, theadjuvant may induce activation of Natural Killer T (NKT) cells. In someembodiments, the adjuvant induces the activation of T_(H)1 cells orT_(H)17 cells or T_(H)2 cells. In other embodiments, the adjuvantinduces the activation of B cells. In yet other embodiments, theadjuvant induces the activation of antigen-presenting cells. Thesecategories are not mutually exclusive; in some cases, an adjuvantactivates more than one type of cell.

In certain embodiments, an adjuvant is a substance that increases thenumbers or activity of antigen presenting cells such as dendritic cells.In certain embodiments, an adjuvant promotes the maturation of antigenpresenting cells such as dendritic cells. In some embodiments, theadjuvant is or comprises a saponin. Typically, the saponin is atriterpene glycoside, such as those isolated from the bark of theQuillaja saponaria tree. A saponin extract from a biological source canbe further fractionated (e.g., by chromatography) to isolate theportions of the extract with the best adjuvant activity and withacceptable toxicity. Typical fractions of extract from Quillajasaponaria tree used as adjuvants are known as fractions A and C. Anexemplary saponin adjuvant is QS-21, which is available from Antigenics.QS-21 is an oligosaccharide-conjugated small molecule. Optionally, QS-21may be admixed with a lipid such as 3D-MPL or cholesterol.

A particular form of saponins that may be used in vaccine formulationsdescribed herein is immunostimulating complexes (ISCOMs). ISCOMs are anart-recognized class of adjuvants, that generally comprise Quillajasaponin fractions and lipids (e.g., cholesterol and phospholipids suchas phosphatidyl choline). In certain embodiments, an ISCOM is assembledtogether with a polypeptide or nucleic acid of interest. However,different saponin fractions may be used in different ratios. Inaddition, the different saponin fractions may either exist together inthe same particles or have substantially only one fraction per particle(such that the indicated ratio of fractions A and C are generated bymixing together particles with the different fractions). In thiscontext, “substantially” refers to less than 20%, 15%, 10%, 5%, 4%, 3%,2% or even 1%. Such adjuvants may comprise fraction A and fraction Cmixed into a ratio of 70-95 A: 30-5 C, such as 70 A:30 C to 75 A:25 C,75 A:25 C to 80 A:20 C, 80 A:20 C to 85 A:15 C, 85 A:15 C to 90 A:10 C,90 A:10 C to 95 A:5 C, or 95 A:5 C to 99 A:1 C. ISCOMatrix, produced byCSL, and AbISCO 100 and 300, produced by Isconova, are ISCOM matricescomprising saponin, cholesterol and phospholipid (lipids from cellmembranes), which form cage-like structures typically 40-50 nm indiameter. Posintro, produced by Nordic Vaccines, is an ISCOM matrixwhere the immunogen is bound to the particle by a multitude of differentmechanisms, e.g. electrostatic interaction by charge modification,incorporation of chelating groups or direct binding.

In some embodiments, the adjuvant is a TLR ligand. TLRs are proteinsthat may be found on leukocyte membranes, and recognize foreign antigens(including microbial antigens). An exemplary TLR ligand is IC-31, whichis available from Intercell. IC31 comprises an antimicrobial peptide,KLK, and an immunostimulatory oligodeoxynucleotide, ODN1a. IC31 has TLR9agonist activity. Another example is CpG-containing DNA, and differentvarieties of CpG-containing DNA are available from Prizer (Coley):VaxImmune is CpG 7909 (a (CpG)-containing oligodeoxy-nucleotide), andActilon is TLR9 agonist, CpG 10101 (a (CpG)-containingoligodeoxy-nucleotide).

In some embodiments, the adjuvant is a nanoemulsion. One exemplarynanoemulsion adjuvant is Nanostat Vaccine, produced by Nanobio. Thisnanoemulsion is a high-energy, oil-in-water emulsion. This nanoemulsiontypically has a size of 150-400 nanometers, and includes surfactants toprovide stability. More information about Nanostat can be found in U.S.Pat. Nos. 6,015,832, 6,506,803, 6,559,189, 6,635,676, and 7,314,624.

Adjuvants may be covalently bound to antigens (e.g., the polypeptidesdescribed above). In some embodiments, the adjuvant may be a proteinwhich induces inflammatory responses through activation ofantigen-presenting cells (APCs). In some embodiments, one or more ofthese proteins can be recombinantly fused with an antigen of choice,such that the resultant fusion molecule promotes dendritic cellmaturation, activates dendritic cells to produce cytokines andchemokines, and ultimately, enhances presentation of the antigen to Tcells and initiation of T cell responses (see Wu et al., Cancer Res2005; 65(11), pp 4947-4954). Other exemplary adjuvants that may becovalently bound to antigens comprise polysaccharides, syntheticpeptides, lipopeptides, and nucleic acids.

The adjuvant can be used alone or in combination of two or more kinds.Adjuvants may be directly conjugated to antigens. Adjuvants may also becombined to increase the magnitude of the immune response to theantigen. Typically, the same adjuvant or mixture of adjuvants is presentin each dose of a vaccine. Optionally, however, an adjuvant may beadministered with the first dose of vaccine and not with subsequentdoses (i.e. booster shots). Alternatively, a strong adjuvant may beadministered with the first dose of vaccine and a weaker adjuvant orlower dose of the strong adjuvant may be administered with subsequentdoses. The adjuvant can be administered before the administration of theantigen, concurrent with the administration of the antigen or after theadministration of the antigen to a subject (sometimes within 1, 2, 6, or12 hours, and sometimes within 1, 2, or 5 days). Certain adjuvants areappropriate for human patients, non-human animals, or both.

2. Additional Components of Vaccines and Pharmaceutical Compositions

In addition to the antigens and the adjuvants described above, a vaccineformulation or pharmaceutical composition may include one or moreadditional components.

In certain embodiments, the vaccine formulation or pharmaceuticalcomposition may include one or more stabilizers such as sugars (such assucrose, glucose, or fructose), phosphate (such as sodium phosphatedibasic, potassium phosphate monobasic, dibasic potassium phosphate, ormonosodium phosphate), glutamate (such as monosodium L-glutamate),gelatin (such as processed gelatin, hydrolyzed gelatin, or porcinegelatin), amino acids (such as arginine, asparagine, histidine,L-histidine, alanine, valine, leucine, isoleucine, serine, threonine,lysine, phenylalanine, tyrosine, and the alkyl esters thereof), inosine,or sodium borate.

In certain embodiments, the vaccine formulation or pharmaceuticalcomposition includes one or more buffers such as a mixture of sodiumbicarbonate and ascorbic acid. In some embodiments, the vaccineformulation may be administered in saline, such as phosphate bufferedsaline (PBS), or distilled water.

In certain embodiments, the vaccine formulation or pharmaceuticalcomposition includes one or more surfactants such as polysorbate 80(Tween 80), Triton X-100, Polyethylene glycol tert-octylphenyl ethert-Octylphenoxypolyethoxyethanol4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol (TRITON X-100);Polyoxyethylenesorbitan monolaurate Polyethylene glycol sorbitanmonolaurate (TWEEN 20); and 4-(1,1,3,3-Tetramethylbutyl)phenol polymerwith formaldehyde and oxirane (TYLOXAPOL). A surfactant can be ionic ornonionic.

In certain embodiments, the vaccine formulation or pharmaceuticalcomposition includes one or more salts such as sodium chloride, ammoniumchloride, calcium chloride, or potassium chloride.

In certain embodiments, a preservative is included in the vaccine. Inother embodiments, no preservative is used. A preservative is most oftenused in multi-dose vaccine vials, and is less often needed insingle-dose vaccine vials. In certain embodiments, the preservative is2-phenoxyethanol, methyl and propyl parabens, benzyl alcohol, and/orsorbic acid.

In certain embodiments, the vaccine formulation or pharmaceuticalcomposition is a controlled release formulation.

E. DNA Vaccines

In certain aspects, the vaccine comprises one of the nucleic acidsdisclosed herein. When a nucleic acid vaccine is administered to apatient, the corresponding gene product (such as a desired antigen) isproduced in the patient's body. In some embodiments, nucleic acidvaccine vectors that include optimized recombinant polynucleotides canbe delivered to a mammal (including humans) to induce a therapeutic orprophylactic immune response. The nucleic acid may be, for example, DNA,RNA, or a synthetic nucleic acid. The nucleic acid may be singlestranded or double stranded.

Nucleic acid vaccine vectors (e.g., adenoviruses, liposomes,papillomaviruses, retroviruses, etc.) can be administered directly tothe mammal for transduction of cells in vivo. The nucleic acid vaccinescan be formulated as pharmaceutical compositions for administration inany suitable manner, including parenteral administration.

In determining the effective amount of the vector to be administered inthe treatment or prophylaxis of an infection or other condition, thephysician evaluates vector toxicities, progression of the disease, andthe production of anti-vector antibodies, if any. Often, the doseequivalent of a naked nucleic acid from a vector is from about 1 μg to 1mg for a typical 70 kilogram patient, and doses of vectors used todeliver the nucleic acid are calculated to yield an equivalent amount oftherapeutic nucleic acid. Administration can be accomplished via singleor divided doses. The toxicity and therapeutic efficacy of the nucleicacid vaccine vectors can be determined using standard pharmaceuticalprocedures in cell cultures or experimental animals.

A nucleic acid vaccine can contain DNA, RNA, a modified nucleic acid, ora combination thereof. In some embodiments, the vaccine comprises one ormore cloning or expression vectors; for instance, the vaccine maycomprise a plurality of expression vectors each capable of autonomousexpression of a nucleotide coding region in a mammalian cell to produceat least one immunogenic polypeptide. An expression vector oftenincludes a eukaryotic promoter sequence, such as the nucleotide sequenceof a strong eukaryotic promoter, operably linked to one or more codingregions. The compositions and methods herein may involve the use of anyparticular eukaryotic promoter, and a wide variety are known; such as aCMV or RSV promoter. The promoter can be, but need not be, heterologouswith respect to the host cell. The promoter used may be a constitutivepromoter.

A vector useful in the present compositions and methods can be circularor linear, single-stranded or double stranded and can be a plasmid,cosmid, or episome. In a suitable embodiment, each nucleotide codingregion is on a separate vector; however, it is to be understood that oneor more coding regions can be present on a single vector, and thesecoding regions can be under the control of a single or multiplepromoters.

Numerous plasmids may be used for the production of nucleic acidvaccines. Suitable embodiments of the nucleic acid vaccine employconstructs using the plasmids VR1012 (Vical Inc., San Diego Calif.),pCMVI.UBF3/2 (S. Johnston, University of Texas) or pcDNA3.1 (InVitrogenCorporation, Carlsbad, Calif.) as the vector. In addition, the vectorconstruct can contain immunostimulatory sequences (ISS), such asunmethylated dCpG motifs, that stimulate the animal's immune system. Thenucleic acid vaccine can also encode a fusion product containing theimmunogenic polypeptide. Plasmid DNA can also be delivered usingattenuated bacteria as delivery system, a method that is suitable forDNA vaccines that are administered orally. Bacteria are transformed withan independently replicating plasmid, which becomes released into thehost cell cytoplasm following the death of the attenuated bacterium inthe host cell.

An alternative approach to delivering the nucleic acid to an animalinvolves the use of a viral or bacterial vector. Examples of suitableviral vectors include adenovirus, polio virus, pox viruses such asalphaviruses, vaccinia, canary pox, and fowl pox, herpes viruses,including catfish herpes virus, adenovirus-associated vector, andretroviruses. Virus-like vectors include virosomes and virus-likeparticles. Exemplary bacterial vectors include attenuated forms ofSalmonella, Shigella, Edwardsiella ictaluri, Yersinia ruckerii, andListeria monocytogenes. In some embodiments, the nucleic acid is avector, such as a plasmid, that is capable of autologous expression ofthe nucleotide sequence encoding the immunogenic polypeptide.

F. Use of Vaccines

The vaccines described herein may be used for prophylactic and/ortherapeutic treatment of herpes, including HSV-1 and particularly HSV-2.The subject receiving the vaccination may be a male or a female, and maybe a child or adult. In some embodiments, the subject being treated is ahuman. In other embodiments, the subject is a non-human animal.

1. Prophylactic Use

In prophylactic embodiments, the HSV-2 vaccine is administered to asubject to induce an immune response that can help protect against theestablishment of HSV-2.

In some embodiments, the vaccine compositions of the invention conferprotective immunity, allowing a vaccinated individual to exhibit delayedonset of symptoms or reduced severity of symptoms (e.g., reduced numberof lesions at the onset of infection), as the result of his/her exposureto the vaccine (e.g., a memory response). In certain embodiments, thereduction in severity of symptoms is at least 25%, 40%, 50%, 60%, 70%,80% or even 90%. Some vaccinated individuals may display no symptomsupon contact with HSV-2 or even no infection by HSV-2. Protectiveimmunity is typically achieved by one or more of the followingmechanisms: mucosal, humoral, or cellular immunity. Mucosal immunity isprimarily the result of secretory IgA (sIGA) antibodies on mucosalsurfaces of the respiratory, gastrointestinal, and genitourinary tracts.The sIGA antibodies are generated after a series of events mediated byantigen-processing cells, B and T lymphocytes, that result in sIGAproduction by B lymphocytes on mucosa-lined tissues of the body. Humoralimmunity is typically the result of IgG antibodies and IgM antibodies inserum. For example, the IgG titer can be raised by 1.5-fold, 2-fold,3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, or even 100-fold ormore following administration of a vaccine formulation described herein.Cellular immunity can be achieved through cytotoxic T lymphocytes orthrough delayed-type hypersensitivity that involves macrophages and Tlymphocytes, as well as other mechanisms involving T cells without arequirement for antibodies. In particular, cellular immunity may bemediated by T_(H)1 cells or T_(H)17 cells. Activation of T_(H)1 cellscan be measured by secretion of IFN-γ, relative to the level of IFN-γreleased in response to a polypeptide that does not generate animmunologic response. In certain embodiments, the amount of IFN-γreleased in 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold,50-fold or even 100-fold greater. The primary result of protectiveimmunity is the destruction of HSV-2 viral particles or inhibition ofHSV-2's ability to replicate. In some embodiments, the protectiveimmunity conferred by presentation of antigen before exposure to HSV-2will reduce the likelihood of seroconversion to an HSV-2-positivestatus.

The duration of protective immunity is preferably as long as possible.In certain embodiments, vaccine formulations produce protective immunitylasting six months, one year, two years, five years, ten years, twentyyears or even a lifetime.

2. Therapeutic Use

In therapeutic applications, the vaccine comprising a polypeptide ornucleic acid of the invention may be administered to a patient sufferingfrom HSV-2, in an amount sufficient to treat the patient. Treating thepatient, in this case, may refer to delaying or reducing symptoms ofHSV-2 in an infected individual. In some embodiments, treating thepatient refers to reducing the duration of lesions, reducing the numberof lesions, reducing the duration of symptoms per episode, and/orotherwise reducing the intensity of symptoms per episode. In certainembodiments, the vaccine reduces the duration or severity of mildsymptoms; in some embodiments, the vaccine reduces the duration orseverity of serious symptoms. In some embodiments, the vaccine reducesviral shedding and therefore the transmissibility of HSV-2 from thevaccinated patient. In certain embodiments, the reductions describedabove are at least 25%, 30%, 40%, 50%, 60%, 70%, 80% or even 90%. Incertain embodiments, the reductions described above include the completecessation of symptoms, viral shedding and/or future outbreaks (e.g., byblocking the ability of the virus to establish latency in sensoryganglia).

In therapeutic embodiments, the HSV-2 vaccine is administered to anindividual post-infection. The HSV-2 vaccine may be administered shortlyafter infection, e.g. before symptoms manifest, or may be administeredduring or after manifestation of symptoms. In some embodiments, theHSV-2 may prevent endogenous reactivation of earlier infection. In someembodiments, a postinfection vaccine could be administered to patientsin high-risk groups.

The duration of therapeutic effects of a vaccine formulation disclosedherein is preferably as long as possible. In certain embodiments,vaccine formulations produce therapeutic effects lasting one month, twomonths, three months, six months, one year, two years, five years, tenyears, twenty years or even a lifetime.

3. Assaying Vaccination Efficacy

The efficacy of vaccination with the vaccines disclosed herein may bedetermined in a number of ways.

Vaccine efficacy may be assayed in various model systems. Suitable modelsystems used to study HSV-2 include a guinea pig model and a mousemodel, as described in the examples below. Briefly, the animals arevaccinated and then challenged with HSV-2 or the vaccine is administeredto already-infected animals. The response of the animals to the HSV-2challenge or the vaccine is then compared with control animals, usingone of the measures described above. A similar assay could be used forclinical testing of humans. The treatment and prophylactic effectsdescribed above represent additional ways of determining efficacy of avaccine.

In addition, efficacy may be evaluated by in vitro immunization of naïvehuman peripheral blood mononuclear cells (PBMC), where APCs are exposedto the vaccine and then the APCs are co-cultured with naïve T cells fromthe same donor to evaluate the primary response to immunization in atest tube. An activation of the T-cells by 1.5 fold, 2-fold, 5-fold,10-fold, 20-fold, 50-fold or 100-fold or more relative to activation ofT-cells using APCs not exposed to a vaccine, in certain embodiments, isconsidered an adequate response.

Vaccine efficacy may further be determined by viral neutralizationassays. Briefly, animals are immunized and serum is collected on variousdays post-immunization. Serial dilutions of serum are pre-incubated withvirus during which time antibodies in the serum that are specific forthe virus will bind to it. The virus/serum mixture is then added topermissive cells to determine infectivity by a plaque assay. Ifantibodies in the serum neutralize the virus, there are fewer plaquescompared to the control group.

G. Uses of Pharmaceutical Compositions

1. Defense Against HSV Infection

The pharmaceutical compositions of the present disclosure are designedto elicit an immune response against HSV-2. Compositions describedherein may stimulate an innate immune response, an antibody response ora cell-mediated immune response, or a combination of these responses, inthe subject to which it is administered. In some embodiments, thecomposition stimulates immune cells at the peripheral site of infectionor sensory ganglia, such as neutrophils, macrophages, and NK cells. Thecomposition may stimulate infiltration by macrophages; production ofantiviral compounds, such including nitric oxide, TNF-α, interferons(IFN), and interleukin 12 (IL-12) by neutrophils; and/or stimulation ofNK cells to produce IFN-γ. IL-2, IFN-α and IFN-β production may also betriggered by the polypeptides of the present composition, and arebelieved to aid in controlling infection.

In some embodiments, the composition comprises antigens that stimulateproduction of neutralizing antibodies. Neutralizing antibodies maytarget the glycoproteins of the viral envelope, which mediate theinteraction of virions with host cell and are responsible forattachment, binding, and entry of HSV-2 into cells. Accordingly, anexemplary composition comprises one or more glycoproteins describedabove or encoded by nucleic acids described above. Immunogenic antigensand/or epitopes as described herein may be administered separately, inseries, or in combination with one another.

In some embodiments, the composition elicits a cell-mediated response,which may involve CD4+ T cells, CD8+ T cells and/or production ofantiviral cytokines. The composition may trigger IFN-γ secretion, forexample through the activation of the innate immune response, andmediate CD8+ T cell clearing of the virus. IFN-γ is also secreted byT_(H)1 cells, (T_(H)17 cells?) T_(C) cells, dendritic cells, and NKcells, and the composition may trigger IFN-γ secretion by any of thesecell types. Such activity of CD8+ T cells may be cytolytic, or,alternately, may be regulated by inhibitor molecules on the surface ofthe neurons which prevent neuronal killing. CD4+ and/or CD8+ T cells mayplay a role in maintaining latency of the virus, thus preventingreactivation. In some embodiments, the composition boosts a CD4+ T cellresponse and/or a CD8+ T cell response that prevents reactivation of thevirus from its latent state.

In some embodiments, the composition blocks the ability of HSV to evadethe host immune response, or, alternately, boosts immune responsesnormally evaded by HSV. In some embodiments, the composition inhibitsHSV-2 from shifting the immunological balance towards tolerance of HSVantigens. HSV-2 may mediate tolerance through T_(H)2 cells. First, HSV-2may induce suppressor T cells, such as CD4+ CD25+ cells and Tr1 cellsthat secrete IL-10, a T_(H)2 cytokine. T_(H)2 cytokines downregulatecostimulatory molecules and inhibit the maturation and function ofantigen-presenting dendritic cells. In addition, infection with HSV-2inhibits the maturation and migration of dendritic cells, which areessential for efficient CTL priming. Notably, T_(H)2 cytokines areproduced during recurrence of HSV-2 infection, in contrast to T_(H)1cytokines, which are produced during recurrence-free episodes. Thus, incertain embodiments, the compositions of the invention represssuppressor T cells and/or induce maturation or migration or both ofdendritic cells.

In some embodiments, methods of inducing an immune response againstHSV-2 in a mammal comprise administering the compositions describedabove. The composition may be used to induce an immune response atdifferent time points, such as before exposure to HSV-2, after initialinfection with HSV-2, before or after HSV-2 has established latency,before or after HSV-2 shedding occurs, and/or before or after recurrentoutbreaks occur. In some embodiments, an immune response against HSV-2may be induced at one or more of the timepoints above. The compositionmay induce a T_(H)1 response and/or a T_(H)17 response but not a T_(H)2response, or may activate the responses at the same time or at differenttimes.

In some embodiments, administration of the composition reduces symptomsassociated with initial infection, latency, or recurrent infection withHSV. Such a composition may reduce incidence and/or severity of lesions,sores, pain, irritation, itching, fever, malaise, headache, viralshedding, or prodromes associated with HSV infection or outbreak.

In some embodiments, one or more antibodies to antigens of HSV-2 may beadministered to individuals in order to produce passive immunity.Passive immunity results from the transfer of active humoral immunity inthe form of ready-made antibodies, from one individual to another.Passive immunization may be used when there is a high risk of infectionand insufficient time for the body to develop its own immune response,or to reduce the symptoms of ongoing or immunosuppressive diseases.Adoptive transfer of T cells may provide another method of eliciting animmune response to HSV-2 antigens in patients. In one embodiment,autologous T cells may be expanded on APCs presenting the antigensderived from the polypeptides described above. Subsequently, theexpanded HSV-2-specific T cells are transferred back into the patientfrom which the T cells were derived.

2. Diagnostic Uses

This application provides, inter alia, a rapid, inexpensive, sensitive,and specific method for detection of HSV-2 in patients. In this respectit should be useful to hospitals and physicians examining and treatingpatients with or at risk for HSV-2 infection. As used herein, “patient”refers to an individual (such as a human) that either has an HSV-2infection or has the potential to contract an HSV-2 infection.

In some embodiments, one may use an antibody against one of thepolypeptides described herein, such as those of Table 1 and/or Table 2,to detect HSV-2 in an individual. The instant disclosure also provides amethod of phenotyping biological samples from patients suspected ofhaving a HSV-2 infection that involves: (a) rendering a biologicalsample amenable to immunoassay, if necessary; (b) contacting the samplewith an appropriate HSV-2-specific antibody or antigen-binding portionthereof under conditions that allow for binding of the antibody orantigen-binding portion to an epitope of HSV-2; and (c) determining ifthe sample shows the presence of HSV-2 as compared to a control tissue;where if the test tissue shows the presence of HSV-2, the patient isidentified as likely having a HSV-2 infection.

Alternatively, one may use the polypeptides described above to detectanti-HSV-2 antibodies in an individual. The instant disclosure alsoprovides a method of phenotyping biological samples from patientssuspected of having a HSV-2 infection: (a) rendering a biological sampleamenable to an affinity assay such as ELISA, if necessary; (b)contacting the sample with a HSV-2-specific antigen or portion thereofunder conditions that allow for binding of the antigen to any hostantibodies present in the sample; and (c) determining if the sampleshows the presence of HSV-2 as compared to a control tissue; wherein ifthe test tissue shows the presence of HSV-2, the patient is identifiedas likely having a HSV-2 infection. The aforementioned test may beappropriately adjusted to detect other viral infections, for instance byusing a homolog (from another viral species) of the proteins describedabove, such as in Table 1 and/or Table 2.

A number of methods for measuring antibody-antigen binding are known inthe art, including ELISA (enzyme-linked immunosorbent assay), Westernblotting, competition assay, and spot-blot. The detection step may be,for instance, chemiluminescent, fluorescent, or colorimetric. Onesuitable method for measuring antibody-protein binding is the LuminexxMAP system, where peptides are conjugated to a dye-containingmicrosphere. Certain systems, including the xMAP system, are amenable tomeasuring several different markers in multiplex, and could be used tomeasure levels of antibodies at once. In some embodiments, other systemsare used to assay a plurality of markers in multiplex. For example,profiling may be performed using any of the following systems: antigenmicroarrays, bead microarrays, nanobarcodes particle technology, arrayedproteins from cDNA expression libraries, protein in situ array, proteinarrays of living transformants, universal protein array, lab-on-a-chipmicrofluidics, and peptides on pins. Another type of clinical assay is achemiluminescent assay to detect antibody binding. In some such assays,including the VITROS Eci anti-HCV assay, antibodies are bound to asolid-phase support made up of microparticles in liquid suspension, anda surface fluorometer is used to quantify the enzymatic generation of afluorescent product.

In other embodiments, one may use the polypeptides described above, suchas those of Table 1 and/or Table 2, to detect T cells that are specificto HSV-2. The instant disclosure provides a method of phentoypingbiological samples from patients suspected of having a HSV-2 infection,involving (a) rendering a biological sample amendable to an assay foractivation of T cells, if necessary, (b) contacting the sample with aHSV-2-specific polypeptide or portion thereof under conditions thatallow APCs to process the polypeptide, and (c) determining activation ofthe T cells in response to the HSV-2-specific polypeptide, where anelevated T cell activation relative to an uninfected patient indicatesHSV-2 infection. This diagnostic assay is intended to detect thepresence of HSV-2-specific T cells in any patients, including thosepatients who have been exposed to HSV-2 but have not seroconverted toproduce detectable levels of anti-HSV-2 antibodies.

T cell activation may be measured using many proliferation assays,including cytokine-specific ELISA, cell proliferation measured bytritiated thymidine incorporation or membrane intercolating (PKH-67) orcytoplasmic (CFSE) dyes, ELISPOT, flow cytometry, and bead arrays. Inaddition, one may measure the T cell response in T cell lines or in Tcell hybridomas from mice or humans that are specific for the antigens.Readouts for activated T cells include proliferation, cytokineproduction, or readout of a surrogate enzyme expressed by the hybridomathat is induced when the T cell or T cell hybridoma is activated inresponse to an antigen. For example, activation of a T cell response maybe detected by T cell hybridoma that is engineered to produceβ-galactosidase. β-galactosidase may be detected through the use ofcolorimetric β-galactosidase substrates such as chlorophenyl red β-Dgalactopyranoside (CPRG).

Infection with HSV-2 may be acute or latent. In some embodiments, if thebiological sample shows the presence of HSV-2, one may administer atherapeutically effective amount of the compositions and therapiesdescribed herein to the patient. The biological sample may comprise, forexample, blood, semen, urine, vaginal fluid, mucus, saliva, feces,urine, cerebrospinal fluid, or a tissue sample. In some embodiments, thebiological sample is an organ intended for transplantation. In certainembodiments, before the detection step, the biological sample is subjectto culture conditions that promote the growth of HSV-2.

The diagnostic tests herein may be used to detect HSV-2 in a variety ofsamples, including samples taken from patients and samples obtained fromother sources. For example, the diagnostic tests may be used to detectHSV-2 on objects such as medical instruments. In some embodiments, thetests herein may be performed on samples taken from animals such asagricultural animals (cows, pigs, chickens, goats, horses and the like),companion animals (dogs, cats, birds, and the like), or wild animals. Incertain embodiments, the tests herein may be performed on samples takenfrom cell cultures such as cultures of human cells that produce atherapeutic protein, cultures of bacteria intended to produce a usefulbiological molecule, or cultures of cells grown for research purposes.

The invention also includes a method of determining the location of aHSV-2 infection in a patient comprising: (a) administering apharmaceutical composition comprising a labeled HSV-2 antibody orantigen-binding portion thereof to the patient, (b) detecting the label,and (c) determining if the patient has HSV-2 compared to a control. Incertain embodiments, the method further comprises, if the patient has anHSV-2 infection, administering a therapeutically effective amount of acomposition described herein to the patient. The method may furthercomprise determining the infected cell types and/or volume of the HSV-2in the patient. This method may be used to evaluate the spread of HSV-2in the patient and determine whether a localized therapy is appropriate.

In some embodiments, the polypeptides described herein may be used tomake a prognosis of the course of infection. In some embodiments, T cellor antibody responses specific for the polypeptides herein may bedetected in a sample taken from a patient. If antibodies or T cells arepresent at normal levels, it would indicate that the patient has raisedan effective immune response against the pathogen. If antibodies or Tcells are absent, or present at reduced levels, it would indicate thatthe patient is failing to raise a sufficient response against thepathogen, and a more aggressive treatment would be recommended. In someembodiments, antibody or T cells present at reduced levels refers toresponses that are present at less than 50%, 20%, 10%, 5%, 2%, or 1% thetypical level in a patient with a protective immune response. T cellresponses may be detected by methods known in the art such as T cellproliferation, ELISPOT or ELISA, and antibodies may be detected byaffinity for any of the antigens described herein, using methods knownin the art such as ELISA.

In some embodiments, detection of T cells specific for HSV-2 antigensmay be used to predict the progress and symptoms of HSV-2 infection in apatient. After infection with HSV-2, some patients remain asymptomatic,although the virus may establish latency. Other patients exhibitsymptoms of HSV-2 infection, and may experience recurrent outbreaks. TheHSV-2 antigens found in asymptomatic patients may differ from thoseantigens found in patients who present symptoms and/or recurrentoutbreaks. Accordingly, the detection methods of the present inventionmay be used to distinguish between subgroups within the population ofpatients infected with HSV-2. Subgroups may be further divided intopatients who experience frequent outbreaks and those who infrequently ornever experience outbreaks, or patients who shed high levels of virusand those who shed low levels or do not shed. The categorization ofpatients, based on the presence and levels of T cell responses tocertain HSV-2 antigens but not others, may help health carepractitioners to determine appropriate treatment regimens. Similarly,differences in the magnitude of T cell responses and/or differences inthe combination and levels of cytokines produced by T cells may also beused to predict the progress and symptoms of HSV-2 infection in apatient. Thus, an infected patient whose complement of HSV-2 antigens towhich T cells respond predicts severe symptoms, frequent outbreaks,and/or high levels of viral shedding may require more intensiveantiviral therapy and/or a longer course of therapeutic treatment than apatient whose complement of HSV-2 antigens predicts an asymptomaticinfection.

It will be understood by one of skill in the art that the methods hereinare not limited to detection of HSV-2. Other embodiments include thedetection of related viruses including viruses with proteins homologousto the proteins described above, such as those in Table 1 and/or Table2. Such related viruses include, for example, other members of theHerpesviridae family. Depending on the homology, these related virusesmay also include viruses that are not members of the Herpesviridaefamily.

3. Use in Groups with Increased Risk for Infection by HSV-2

Essentially any individual has a certain risk of infection with HSV-2.However, certain sub-populations have an increased risk of infection. Insome embodiments, patients receiving the composition for HSV-2 areimmunocompromised.

An immunocompromising condition arising from a medical treatment islikely to expose the individual in question to a higher risk ofinfection. It is possible to treat an infection prophylactically in anindividual having the immunocompromised condition before or duringtreatments known to generate such a condition. By prophylacticallytreating with the antigen before or during a treatment known to generatesuch a condition it is possible to prevent a subsequent infection or toreduce the risk of the individual contracting an infection due to theimmunocompromised condition. Should the individual contract aninfection, e.g., following a treatment leading to an immunocompromisedcondition, it is also possible to treat the infection by administeringto the individual an antigen composition.

In certain embodiments, the compositions are administered to children oradult patients. In other embodiments, compositions are appropriate forpregnant women who were infected before becoming pregnant, or who becameinfected during pregnancy, such as to inhibit infection of a fetus orbaby. The compositions may also be administered to neonates and infantswho became infected in utero or during delivery.

H. Doses and Routes of Administration

1. Dosage Amounts and Timing

The amount of antigen in each vaccine dose is selected as an effectiveamount, which induces an prophylactic or therapeutic response, asdescribed above, in either a single dose or over multiple doses.Preferably, the dose is without significant adverse side effects intypical vaccinees. Such amount will vary depending upon which specificantigen is employed. Generally, it is expected that a dose will comprise1-1000 μg of protein, in some instances 2-100 μg, for instance 4-40 μg.An optimal amount for a particular vaccine can be ascertained bystandard studies involving observation of antibody titers, T cellactivation levels, and other responses in subjects. In some embodiments,the appropriate amount of antigen to be delivered will depend on theage, weight, and health (e.g. immunocompromised status) of a subject.When present, typically an adjuvant will be present in amounts from 1μg-250 μg per dose, for example 50-150 μg, 75-125 μg or 100 μg.

In some embodiments, only one dose of the vaccine is administered toachieve the results described above. In other embodiments, following aninitial vaccination, subjects receive one or more boost vaccinations,for a total of two, three, four or five vaccinations. Advantageously,the number is three or fewer. A boost vaccination may be administered,for example, about 1 month, 2 months, 4 months, 6 months, or 12 monthsafter the initial vaccination, such that one vaccination regimeninvolves administration at 0, 0.5-2 and 4-8 months. It may beadvantageous to administer split doses of vaccines which may beadministered by the same or different routes.

The pharmaceutical compositions described herein may take on a varietyof dosage forms. In certain embodiments, the composition is provided insolid or powdered (e.g., lyophilized) form; it also may be provided insolution form. In certain embodiments, a dosage form is provided as adose of lyophilized composition and at least one separate sterilecontainer of diluent.

In some embodiments, the antigen is delivered to a patient at an amountof 1 μmol per dose. In some embodiments, the antigen is delivered at adose ranging from 10 nmol to 100 nmol per dose. The appropriate amountof antigen to be delivered may be determined by one of skill in the art.In some embodiments, the appropriate amount of antigen to be deliveredwill depend on the age, weight, and health (e.g., immunocompromisedstatus) of a subject.

Pharmaceutical compositions disclosed herein are (in some embodiments)administered in amounts sufficient to elicit production of antibodies aspart of an immunogenic response. In some embodiments, the compositionmay be formulated to contain 5 mcg/0.5 mL or an amount ranging from 10mcg/1 mL to 200 mcg/1 mL of an antigen. In other embodiments, thecomposition may comprise a combination of antigens. The plurality ofantigens may each be the same concentration, or may be differentconcentrations.

In some embodiments, the composition will be administered in a doseescalation manner, such that successive administrations of thecomposition contain a higher concentration of composition than previousadministrations. In some embodiments, the composition will beadministered in a manner such that successive administrations of thecomposition contain a lower concentration of composition than previousadministrations.

In therapeutic applications, compositions are administered to a patientsuffering from a disease in an amount sufficient to cure or at leastpartially arrest the disease and its complications.

Therapeutic applications of a composition described herein includereducing transmissibility, slowing disease progression, reducing viralshedding, or eliminating recurrent infections in patients that have beeninfected with HSV-2, such as by 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%or 10% of the levels at which they would occur in individuals who arenot treated with the composition. The composition may also reduce thequantity of HSV-2 shed by infected individuals, inhibit the expressionof proteins required for reactivation of HSV-2 from the latent stage ininfected patients, and/or inhibit replication of HSV-2 in neurons ofinfected patients, such as by 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or10% of the levels at which they would occur in individuals not treatedwith the composition.

In prophylactic embodiments, compositions are administered to a human orother mammal to induce an immune response that can inhibit theestablishment of an infectious disease or other condition. In someembodiments, a composition may partially block the virus fromestablishing latency or reduce the efficiency with which latency isestablished.

In some embodiments, only one dose (administration) of the compositionis given. In other embodiments, the composition is administered inmultiple doses. In various embodiments, the composition is administeredonce, twice, three times, or more than three times. The number of dosesadministered to a subject is dependent upon the antigen, the extent ofthe disease or the expected exposure to the disease, and the response ofa subject to the composition.

In some embodiments, the compositions are administered in combinationwith antimicrobial molecules. Antimicrobial molecules may includeantiviral molecules. Many antiviral molecules are currently known in theart, and target one or more stage of the viral life cycle, includingviral attachment to host cells, release of viral genes and/or enzymesinto the host cell, replication of viral components using host-cellmachinery, assembly of viral components into complete viral particles,and release of viral particles to infect new hosts.

2. Routes of Administration

The vaccine formulations and pharmaceutical compositions herein can bedelivered by administration to an individual, typically by systemicadministration (e.g., intravenous, intraperitoneal, intramuscular,intradermal, subcutaneous, transdermal, subdermal, intracranial,intranasal, mucosal, anal, vaginal, oral, sublingual, buccal route orthey can be inhaled) or they can be administered by topical application.

In some embodiments, the composition may be administered directly to thelikely sites of infection. In female patients, the composition may beapplied topically to mucosal membranes, or delivery vaginally orrectally using devices and methods known in the art. The vaginal andrectal routes of delivery permits extended, continuous or pulseddelivery and administration of composition dosages, and may beadministered either before or after exposure to HSV, depending on theuse of a prophylactic or therapeutic composition. In male patients, thecomposition may be applied topically to the skin or mucosal membranes,or delivered rectally. In both patient populations, the composition mayalso be targeted to the sensory ganglia.

An HSV-2 vaccine or pharmaceutical composition is often administered viathe intramuscular route. Typically, in this route, the vaccine isinjected into an accessible area of muscle tissue. Intramuscularinjections are, in some embodiments, given in the deltoid, vastuslateralis, ventrogluteal or dorsogluteal muscles. The injection istypically given at an approximately 90° angle to the surface of theskin, so the vaccine penetrates the muscle.

An HSV-2 vaccine may also be administered subcutaneously. The injectionis typically given at a 45° angle to the surface of the skin, so thevaccine is administered to the subcutis and not the muscle.

In some embodiments, the HSV-2 vaccine is administered intradermally.Intradermal administration is similar to subcutaneous administration,but the injection is not as deep and the target skin layer is thedermis. The injection is typically given at a 10-15° angle to thesurface of the skin, so the vaccine is delivered just beneath theepidermis.

3. Formulations

The vaccine formulation may be suitable for administration to a humanpatient, and vaccine preparation may conform to USFDA guidelines. Insome embodiments, the vaccine formulation is suitable for administrationto a non-human animal. In some embodiments, the vaccine is substantiallyfree of either endotoxins or exotoxins. Endotoxins include pyrogens,such as lipopolysaccharide (LPS) molecules. The vaccine may also besubstantially free of inactive protein fragments. In some embodiments,the vaccine has lower levels of pyrogens than industrial water, tapwater, or distilled water. Other vaccine components may be purifiedusing methods known in the art, such as ion-exchange chromatography,ultrafiltration, or distillation. In other embodiments, the pyrogens maybe inactivated or destroyed prior to administration to a patient. Rawmaterials for vaccines, such as water, buffers, salts and otherchemicals may also be screened and depyrogenated. All materials in thevaccine may be sterile, and each lot of the vaccine may be tested forsterility. Thus, in certain embodiments the endotoxin levels in thevaccine fall below the levels set by the USFDA, for example 0.2endotoxin (EU)/kg of product for an intrathecal injectable composition;5 EU/kg of product for a non-intrathecal injectable composition, and0.25-0.5 EU/mL for sterile water.

In some embodiments, the vaccine comprising a polypeptide contains lessthan 5%, 2%, 1%, 0.5%, 0.2%, 0.1% of other, undesired unpolypeptides,relative to the amount of desired polypeptides. In some embodiments, thevaccine contains less than 5%, less than 2%, less than 1%, less than0.5%, less than 0.2%, or less than 0.1% DNA and/or RNA.

It is preferred that the vaccine has low or no toxicity, within areasonable risk-benefit ratio.

The formulations suitable for introduction of the pharmaceuticalcomposition vary according to route of administration. Formulationssuitable for parenteral administration, such as, for example, byintraarticular (in the joints), intravenous, intramuscular, intradermal,intraperitoneal, intranasal, and subcutaneous routes, include aqueousand non-aqueous, isotonic sterile injection solutions, which can containantioxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.The formulations can be presented in unit-dose or multi-dose sealedcontainers, such as ampoules and vials.

Injection solutions and suspensions can be prepared from sterilepowders, granules, and tablets of the kind previously described. Cellstransduced by the packaged nucleic acid can also be administeredintravenously or parenterally.

Formulations suitable for oral administration can consist of (a) liquidsolutions, such as an effective amount of the polypeptides or packagednucleic acids suspended in diluents, such as water, saline or PEG 400;(b) capsules, sachets or tablets, each containing a predetermined amountof the active ingredient, as liquids, solids, granules or gelatin; (c)suspensions in an appropriate liquid; and (d) suitable emulsions. Tabletforms can include one or more of lactose, sucrose, mannitol, sorbitol,calcium phosphates, corn starch, potato starch, tragacanth,microcrystalline cellulose, acacia, gelatin, colloidal silicon dioxide,croscarmellose sodium, talc, magnesium stearate, stearic acid, and otherexcipients, colorants, fillers, binders, diluents, buffering agents,moistening agents, preservatives, flavoring agents, dyes, disintegratingagents, and pharmaceutically compatible carriers. Lozenge forms cancomprise the active ingredient in a flavor, usually sucrose and acaciaor tragacanth, as well as pastilles comprising the active ingredient inan inert base, such as gelatin and glycerin or sucrose and acaciaemulsions, gels, and the like containing, in addition to the activeingredient, carriers known in the art. The pharmaceutical compositionscan be encapsulated, e.g., in liposomes, or in a formulation thatprovides for slow release of the active ingredient.

The antigens, alone or in combination with other suitable components,can be made into aerosol formulations (e.g., they can be “nebulized”) tobe administered via inhalation. Aerosol formulations can be placed intopressurized acceptable propellants, such as dichlorodifluoromethane,propane, nitrogen, and the like.

Suitable formulations for vaginal or rectal administration include, forexample, suppositories, which consist of the polypeptides or packagednucleic acids with a suppository base. Suitable suppository basesinclude natural or synthetic triglycerides or paraffin hydrocarbons. Inaddition, it is also possible to use gelatin rectal capsules whichconsist of a combination of the polypeptides or packaged nucleic acidswith a base, including, for example, liquid triglycerides, polyethyleneglycols, and paraffin hydrocarbons. The formulation may be suitable foradministration to a human patient, and the preparation may conform to USFDA guidelines. In some embodiments, the formulation is suitable foradministration to a non-human animal. In some embodiments, thecomposition is substantially free of either endotoxins or exotoxins.Endotoxins may include pyrogens, such as lipopolysaccharide (LPS)molecules. The composition may also be substantially free of inactiveprotein fragments which may cause a fever or other side effects. In someembodiments, the composition contains less than 1%, less than 0.1%, lessthan 0.01%, less than 0.001%, or less than 0.0001% of endotoxins,exotoxins, and/or inactive protein fragments. In some embodiments, thecomposition has lower levels of pyrogens than industrial water, tapwater, or distilled water. Other components may be purified usingmethods known in the art, such as ion-exchange chromatography,ultrafiltration, or distillation. In other embodiments, the pyrogens maybe inactivated or destroyed prior to administration to a patient. Rawmaterials for compositions, such as water, buffers, salts and otherchemicals may also be screened and depyrogenated. All materials in thecomposition may be sterile, and each lot of the composition may betested for sterility. Thus, in certain embodiments the endotoxin levelsin the composition fall below the levels set by the USFDA: 0.2 endotoxin(EU)/kg of product for an intrathecal injectable composition; 5 EU/kg ofproduct for a non-intrathecal injectable composition, and 0.25-0.5 EU/mLfor sterile water.

In certain embodiments, the preparation comprises less than 50%, 20%,10%, or 5% (by dry weight) contaminating protein. In certainembodiments, the desired molecule is present in the substantial absenceof other biological macromolecules, such as other proteins (particularlyother proteins which may substantially mask, diminish, confuse or alterthe characteristics of the component proteins either as purifiedpreparations or in their function in the subject reconstituted mixture).In certain embodiments, at least 80%, 90%, 95%, 99%, or 99.8% (by dryweight) of biological macromolecules of the same type present (butwater, buffers, and other small molecules, especially molecules having amolecular weight of less than 5000, can be present).

It is preferred that the composition has low or no toxicity, within areasonable risk-benefit ratio. In certain embodiments, the compositioncomprises ingredients at concentrations that are less than LD₅₀measurements for the animal being treated with the composition. LD₅₀measurements may be obtained in mice or other experimental modelsystems, and extrapolated to humans and other animals. Methods forestimating the LD₅₀ of compounds in humans and other animals arewell-known in the art. A composition, and any component within it, mighthave an LD₅₀ value in rats of greater than 100 g/kg, greater than 50g/kg, greater than 20 g/kg, greater than 10 g/kg, greater than 5 g/kg,greater than 2 g/kg, greater than 1 g/kg, greater than 500 mg/kg,greater than 200 mg/kg, greater than 100 mg/kg, greater than 50 mg/kg,greater than 20 mg/kg, or greater than 10 mg/kg. In some embodiments,the therapeutic index of the composition (measured as the toxic dose for50% of the population (TD₅₀) divided by the minimum effective dose for50% of the population (ED₅₀)), is greater than 1, greater than 10, orgreater than 100.

I. Preparation and Storage of Vaccines Formulations and ImmunogenicCompositions

The HSV-2 vaccines described herein may be produced using a variety oftechniques. For example, a polypeptide may be produced using recombinantDNA technology in a suitable host cell. A suitable host cell may bebacterial, yeast, mammalian, or other type of cell. The host cell may bemodified to express an exogenous copy of one of the relevant polypeptidegenes. Typically, the gene is operably linked to appropriate regulatorysequences such as a strong promoter and a polyadenylation sequence. Insome embodiments, the promoter is inducible or repressible. Otherregulatory sequences may provide for secretion or excretion of thepolypeptide of interest or retention of the polypeptide of interest inthe cytoplasm or in the membrane, depending on how one wishes to purifythe polypeptide. The gene may be present on an extrachromosomal plasmid,or may be integrated into the host genome. One of skill in the art willrecognize that it is not necessary to use a nucleic acid 100% identicalto the naturally-occurring sequence. Rather, some alterations to thesesequences are tolerated and may be desirable. For instance, the nucleicacid may be altered to take advantage of the degeneracy of the geneticcode such that the encoded polypeptide remains the same. In someembodiments, the gene is codon-optimized to improve expression in aparticular host. The nucleic acid may be produced, for example, by PCRor by chemical synthesis.

Once a recombinant cell line has been produced, a polypeptide may beisolated from it. The isolation may be accomplished, for example, byaffinity purification techniques or by physical separation techniques(e.g., a size column).

In a further aspect of the present disclosure, there is provided amethod of manufacture comprising mixing one or more polypeptides or animmonogenic fragment or variant thereof with a carrier and/or anadjuvant. In some embodiments, the adjuvant is one that stimulates aT_(H)1 cell response.

In some embodiments, antigens for inclusion in compositions of theinvention may be produced in cell culture. One method comprisesproviding one or more mammalian expression vectors and cloningnucleotides encoding two or more polypeptides selected from polypeptideshaving an amino acid sequence of any one of SEQ ID NOS: 1-38, thenexpressing and isolating the polypeptides.

The immunogenic polypeptides described herein, and nucleic acidcompositions that express the polypeptides, can be packaged in packs,dispenser devices, and kits for administering nucleic acid compositionsto a mammal. For example, packs or dispenser devices that contain one ormore unit dosage forms are provided. Typically, instructions foradministration of the compounds will be provided with the packaging,along with a suitable indication on the label that the compound issuitable for treatment of an indicated condition, such as thosedisclosed herein.

V. EXAMPLES Example 1 Identification of HSV-2 Antigens

A library of HSV-2 antigens (from HSV-2 Strain G, Lot #7C0013, fromAdvanced Biotechnologies Inc, Maryland) was prepared and screened withperipheral blood mononuclear cells (PBMC) from human donors. Briefly, alibrary of HSV antigens was expressed by bacteria and mixed with antigenpresenting cells (APCs). The APCs, in turn, presented HSV-derivedpeptides to lymphocytes that had been isolated from human patientsinfected with HSV-2. The patients belonged to several populations, asdescribed below. Lymphocyte responses from each population were comparedfor reactivity to each expressed protein, and the screen detectedantigens that induced reactive lymphocytes with greater frequency in onepatient population as compared to the others. Infected but asymptomatic,and exposed but seronegative patients may activate protective immuneresponses that patients who experience frequent outbreaks do not; inparticular, exposed but seronegative patients are presumed to havemounted sterilizing immunity to HSV-2 infection. It is believed that aunique set of polypeptides will activate lymphocytes from these patientpopulations.

The release of IFN-γ from CD4⁺ T cells and CD8⁺ T cells from eachpopulation was measured by ELISA following exposure to candidateantigens. Antigens were selected on the basis of the fold increase ofIFN-γ released, relative to the level of IFN-γ released by frequentrecurrers who experience more than four outbreaks per year.

A. Identification of Antigens Encoded by UL10, UL19, UL40, US4, US6, RS1(RS1.1, RS1.2, RS1.3), UL 36 (UL36.3, UL36.4, UL36.5), UL32, and RL2

Lymphocytes were isolated from patients belonging to severalpopulations: asymptomatic (n=40), exposed (n=40), frequent recurrers whoexperience 4 or more outbreaks per year (n=43), less-frequent recurrerswho experience less than 4 outbreaks per year (n=19), naïve (n=10), andHSV-2⁻/HSV-1⁺ (n=10). Table 3 shows the frequency analysis for thirteenHSV-2 antigens encoded by UL10, UL19, UL40, US4, US6, RS1 (RS1.1, RS1.2,RS1.3), UL36 (UL36.3, UL 36.4, UL36.5), UL32, and RL2 in the exposedpatient cohort compared to recurrers with 2 or more outbreaks per year.

TABLE 3 Frequency analysis for antigens encoded by UL10, UL19, UL40,US4, US6, RS1 (RS1.1, RS1.2, RS1.3), UL36 (UL36.3, UL36.4, UL36.5), UL32and RL2 Frequency Analysis (HSV-1/HSV-2 seronegative) Protein % responsefrom fold increase over HSV-2 Gene Name exposed donors recurrer responseUL10 gM 23% 1.4 UL19 VP5 — — UL40 ribonucleotide 36% 3.0 reductase Us4gG 24% 1.6 Us6 gD 27% 1.9 RS1 ICP4 RS1.1 54% 3.0 RS1.2 46% 2.3 RS1.3 23%1.2 UL36 Major tegument UL36.3 protein 46% 2.3 UL36.4 46% 4.2 UL36.5 31%1.9 UL32 DNA cleavage & — — packaging proteiin RL2 ICP0 45% 1.6B. Identification of Antigens Encoded by UL1, UL49.5, and UL54

Lymphocytes were isolated from patients belonging to severalpopulations: asymptomatic (n=40), exposed (n=40), frequent recurrers whoexperience 4 or more outbreaks per year (n=43), less-frequent recurrerswho experience less than 4 outbreaks per year (n=19), naïve (n=10), andHSV-2⁻/HSV-1⁺ (n=10).

Table 4 shows the frequency analysis for three HSV-2 antigens encoded byUL1, UL49.5 and UL54, in the exposed patient cohort compared torecurrers with 2 or more outbreaks per year.

TABLE 4 Frequency analysis for antigens encoded by UL1, UL49.5, and UL54Frequency Analysis (HSV-1/HSV-2 seronegative) Protein % response fromfold increase over HSV-2 Gene Name exposed donors recurrer response UL1gL2 64% 2.7 UL49.5 (virion p) 37% 2.1 UL54 ICP27 22% 5.8C. Identification of Antigens Encoded by RL1, UL2, and UL11

Lymphocytes were isolated from patients belonging to severalpopulations: asymptomatic (n=40), exposed (n=40), frequent recurrers whoexperience 4 or more outbreaks per year (n=43), less-frequent recurrerswho experience less than 4 outbreaks per year (n=19), naïve (n=10), andHSV-2⁻/HSV-1⁺ (n=10).

Table 5 shows the frequency analysis for three HSV-2 antigens encoded byRL1, UL2, and UL11 in the exposed patient cohort compared to recurrerswith 2 or more outbreaks per year.

TABLE 5 Frequency analysis for HSV-2 antigens encoded by RL1, UL2, andUL11 Frequency Analysis (HSV-l/HSV-2 seronegative) Protein % responsefrom fold increase over HSV-2 Gene Name exposed donors recurrer responseRL1 ICP34.5 45% 1.3 UL2 DNA 23% 1.4 glycosylase UL11 tegument 21% <1.0protein

Example 2 In Vivo Data

-   A. [Protocol A] Guinea Pig Therapeutic Vaccination Protocol

Female Hartley guinea pigs were challenged intravaginally with HSV-2strain MS at 5×10⁵ pfu to establish a genital tract infection. Animalswere monitored for infection by vaginal swab on day 1 post-infection,and acute disease between days 3 and 14 post-infection. On day 14, afterresolution of primary disease, the animals were randomized into groupsof 12 and immunized subcutaneously with antigen (HSV-2 polypeptide at 15μg dose) plus adjuvant (50 μg dose of an ISCOM matrix with a 91:9mixture of Quillaja saponin fractions A and C). Each group received atotal of 3 vaccinations, on days 14, 21, and 34 post-infection. Genitalswabs were collected during the vaccination period to monitor viralshedding, and daily observations were recorded. Symptoms were scored ona scale from 0 to 4 based upon severity, 0=no symptoms; 1=redness orswelling; 2=a few small vesicles; 3=several large vesicles; 4=severallarge vesicles with maceration. In addition, animals with lesionsintermediate in severity between the above scores were given a score of0.5, 1.5, 2.5, or 3.5.

1. Results of Therapeutic Vaccination Studies with ICP4.2, gD2ΔTMR, andgD2

The results of the studies are presented below in Tables 6-10. The IgGtiter was determined at day 41 post-infection and 7 days after thirdimmunization using an average of 4 out of the 12 animals in each group.The mean recurrent lesion scores and mean lesion days were eachdetermined from day 15 to day 63 post-infection. The lesion scoresrepresent total lesions for each group from day 15 to 60 and then a meanwas calculated. Mean lesion days represent the mean number of dayspost-infection that immunized or non-immunized animals had herpeticlesions present. Vaginal-swab samples were collected from all animalsfor 12 days between days 20-59 post-infection and stored at −80° C.until assayed for virus shedding titers by quantitative real-time PCR.

TABLE 6 Results of therapeutic vaccination studies with ICP4.2 (SEQ IDNO: 2): lesions Mean gD2 Recurrent % Mean % Groups IgG Lesion Reduc-Lesion Reduc- N = 12 Dose Titer Score tion Days tion Phosphate- — 1:2638.1 — 9.0 — Buffered Saline adjuvant 50 μg × 1:331 7.1 14 8.5 6 only 3ICP4.2 + 15 μg ×  1:1079 4.3 47 5.1 44 adjuvant 3

TABLE 7 Results of therapeutic vaccination studies with ICP4.2 (SEQ IDNO: 2): viral shedding No. of animals Mean number with no of daysdetectable viral viral shedding % Reduc- P Groups shedding/totaldetected ± SEM tion value* Phosphate- 0/11 4.5 ± 0.8 — — Buffered SalineAdjuvant only 0/12 4.4 ± 0.7 2 0.971 ICP4.2 + 5/11 1.5 ± 0.5 67 0.004adjuvant

TABLE 8 Results of therapeutic vaccination studies with gD2ΔTMR (SEQ IDNO: 4): lesions Mean Recurrent Mean Lesion Groups Lesion Score %Reduction Days % Reduction Adjuvant only 8.7 — 11.7 — gD2ΔTMR 5.7 34 8.626

TABLE 9 Results of therapeutic vaccination studies with gD2 (SEQ ID NO:5): lesions Mean gD2 Recurrent % Mean % Groups IgG Lesion Reduc- LesionReduc- N = 12 Dose Titer Score tion Days tion Phosphate- — 1:263 8.1 —9.0 — Buffered Saline Adjuvant 50 μg × 1:331 7.1 14 8.5 6 only 3 gD2 +15 μg × >1:6400 4.0 51 5.0 45 adjuvant 3 (p = 0.04)

TABLE 10 Results of therapeutic vaccination studies with gD2 (SEQ ID NO:5): viral shedding No. of animals/total Mean number with no of daysdetectable viral shedding % Reduc- P Groups viral shedding detected ±SEM tion value* Phosphate- 0/11 4.5 ± 0.8 — — Buffered Saline Adjuvantonly 0/12 4.4 ± 0.7 2 0.971 gD2 + adjuvant 4/12 2.4 ± 0.6 47 0.047B. [Protocol B] Murine Prophylactic Vaccination Protocol

Female C57BL/6 mice from 6 to 8 weeks of age were immunizedsubcutaneously with antigen (HSV-2 polypeptide) plus adjuvant (12 μgdose of an ISCOM matrix with a 82:18 mixture of Quillaja saponinfractions A and C) on day 0 and day 9. On day 11, estrous cycles weresynchronized with depo provera and then the mice were challenged on day16 via intravaginal deposition of 10 times the LD₅₀ of HSV-2 strain 333while under anaesthesia. All animals were monitored for morbidity(clinical score) and mortality, and body weights and vaginal swabs werecollected between days 17 and 28 post-infection. Clinical scores wererecorded using the following scale: 0=no symptoms, 1=vaginal erythema,2=vaginal erythema and edema, 3=vaginal herpetic lesions, 4=unilateralparalysis or severe genital ulceration, and 5=bilateral paralysis ordeath.

1. Results of Murine Prophylactic Vaccination Studies with ICP4.2, VP5,gD2ΔTMR and gD2ΔTMR and ICP4.2

In the experimental group, mice were immunized subcutaneously witheither 5 μg or 10 μg of antigen plus adjuvant (12 μg dose of an ISCOMmatrix with a 82:18 mixture of Quillaja saponin fractions A and C) onday 0 and day 9. Control animals received phosphate buffered saline(PBS) only, or adjuvant only.

Mice receiving PBS only or adjuvant only all died by day 9post-challenge (no survivors). In contrast, mice receiving antigenlargely survived to day 9, and 20-75% survived to day 12 post-challenge.The severity of disease symptoms (genital and neurological disease) werealso scored at either day 9 or 10 post-challenge. Mice immunized withICP4.2, VP5, gD2ΔTMR, or gD2ΔTMR and ICP4.2 with ISCOM adjuvant showed asignificant decrease in disease symptoms compared to the PBS only oradjuvant only groups.

TABLE 11 Results of murine prophylactic vaccination studies Mean Disease% Score % Reduc- P Survival Groups Day 10 tion value* Day 12 PBSonly/adjuvant only 5.00/4.81 — — % ICP4.2 3.6 28 — 2.0% VP5 + adjuvant3.13 35 .146 3.8% gD2ΔTMR + adjuvant 1.44 70 .023 7.5% gD2ΔTMR +ICP4.2 + 0.75 84 .020 8.8% adjuvant *Student's t testC. [Protocol C] Guinea Pig Prophylactic Vaccination ProtocolFemale Hartley guinea pigs from 250-350 grams (weight) were immunizedsubcutaneously with 15 μg of antigen plus adjuvant (50 μg dose of anISCOM matrix with a 91:9 mixture of Quillaja saponin fractions A and C)on day 0 and day 14-21. Sera were collected by toenail clip 2-3 weeksafter the boost and then the guinea pigs were challenged viaintravaginal deposition of 5×10⁵ PFU of HSV-2 strain MS. Vaginal-swabsamples were collected from all animals on days 30 and 32 and stored at−80° C. until assayed for virus titers by quantitative real-time PCR.Guinea pigs were evaluated daily (day 1-14), and primary genital skindisease was quantified using a lesion severity score scale from 1-4.Numerical scores were assigned to specific disease signs as follows: 0,no disease; 1, redness or swelling; 2, a few small vesicles; 3, severallarge vesicles; 4, several large vesicles with maceration. At the end ofthe study, the guinea pigs were euthanized, and the dorsal root ganglia(DRG) were harvested, stored at −80° C. until they were processed forquantitative real-time PCR analysis.

TABLE 12 Results of guinea pig prophylactic vaccination studies withgD2ΔTMR and VP5 Total Copies mean HSV-2 Viral titer, acute % DNA/1 μg %PFU/ml lesion Reduc- DRG Reduc- Groups Day 2 score tion DNA tionAdjuvant only 2.3 × 10⁶ 22.6 — 959 — gD2ΔTMR + 1.7 × 10⁶ 7.7 66% 274 71%Adjuvant VP5 + adjuvant 5.9 × 10⁵ 18.2 17% 283 70%D. [Protocol D] Immunogenicity Assay I (Standard)

Mice were immunized subcutaneously in the scruff of the neck with a 100μl injection of 5 μg antigen plus adjuvant (12 μg dose of an ISCOMmatrix with a 82:18 mixture of Quillaja saponin fractions A and C) insaline. The mice received one or two injections, 7 days apart. Analysisof the immunogenicity of the injection occurred 7 days after the finalinjection.

The immunogenicity assay was an ex vivo IFN-γ ELISPOT. CD4⁺ and CD8⁺ Tcells were enriched from the spleen and analyzed separately. For theELISPOT assay, membrane plates were prepared by coating them overnightwith capture antibody and subsequently blocked by supplemented mediumfor a minimum of 2 hours at 37° C. The mice were euthanized and theirspleens harvested. The T cells were then prepared by sorting thesplenocytes for CD4⁺ and CD8⁺ T cells using magnetic beads. The blockingsolution was washed out from ELISPOT plates and the T cells were platedout onto the blocked plates. The plates were returned to the incubatorto allow the T cells to settle. APCs were prepared by pulsing naïveT-depleted splenocytes with antigen for 2 hours at 37° C. For CD4⁺ELISPOTs, APCs were pulsed with whole protein. For CD8⁺ ELISPOTs, APCswere pulsed with E. coli expressing protein plus cLLO. A medium controlwas APCs incubated for 2 hours at 37° C. with no additional antigen. Thepulsed APCs were irradiated, washed and adjusted to 2×10⁶ cells/ml. TheAPCs were added to appropriate wells of plates containing T cells. Thenphorbol myristate acetate (PMA) and ionomycin were added to controlwells as a positive control. The plates were allowed to incubate for 18hours at 37° C. under 5% CO₂. The plates were then developed using asecondary biotinylated antibody, horseradish peroxidase (HRP) and3-amino-9-ethylcarbazole (AEC) substrate.

1. Results of Immunogenicity Assay I with ICP4.2

The immunogenicity assay I showed a robust immunogenic response for boththe one and two injection regimens with ICP4.2. For the one injectionregimen, the number of IFN-γ spots per 200,000 T cells were 8 and 101for CD4⁺ and CD8⁺ cells, respectively. For the two injection regimen,there were 50 and 70 spots, respectively. In contrast, less than 15spots were observed for media or adjuvant alone in either CD4⁺ or CD8⁺cells.

2. Results of Immunogenicity Assay I with gD2 ΔTMR and gD2

Results of immunogenicity assay I are shown in FIGS. 1A and B. RobustCD4⁺ and CD8⁺ T cell responses were obtained for both full-length gD2and for gD2ΔTMR. In contrast, gD2 antigen truncated immediately upstreamof the transmembrane domain (denoted 306t in FIG. 1) showedsignificantly reduced responses.

E. [Protocol E] Immunogenicity Assay II (Rapid)

Recombinant E. coli from Genocea's proprietary library of HSV-2 orfeomewere induced to express gL2 or fragments of ICP4 protein (ICP4.2, andpolypeptides encoded by RS1.1, RS1.3.1 and RS 1.3.2). The protein wasretained within bacterial cells. The bacteria were then fixed with PFA,washed extensively with PBS and stored at −80 C until used forimmunization.

Three mice per group were immunized with 1×10⁸ bacteria in PBS per mouseby intraperitoneal injection. Mice received 1-2 additional boosters at 1week intervals. Seven days after last boost, sera were collected andanalyzed in an HSV-2 neutralization assay. Five-fold serial dilutionswere prepared for plasma or serum samples in a 96-well round-bottomplate, followed by the addition of 50 PFUs HSV-2 (strain 333) to eachwell. The plates were covered and incubated at 37° C. for 1 hour. 200 μlof virus-serum dilution was transferred in duplicate to Vero cells grownin a 48-well tissue culture plate and incubated for 1 hour at 37° C. 300μl of DMEM containing 2% FBS was then added to each well and the plateswere incubated for 48 hours at 37° C. To visualize virus plaques theplates were stained with crystal violet.

TABLE 13 Results of HSV-2 neutralization assay with gL2, ICP4.2, andpolypeptides encoded by RS1.1, RS1.3.1 and RS1.3.2 Immunogen HSV-2Neutralization IgG Titer* E coli//gL2  1:50 E coli//RS1.1 <1:20 Ecoli//ICP4.2 <1:20 E. coli/RS 1.3.1  1:100 E. coli//RS1.3.2 <1:20Positive control    1:2500 (DL11 Mab) Negative control <1:20 (Naïvemouse serum) *Serum dilution that inhibits 50% of virus controlF. [Protocol F] Immunogenicity Assay III (Overlapping Peptide Pools)

Mice were immunized with 2 μg/mouse of pooled, overlapping peptides(OLP) spanning the entire sequence of gL2, ICP4, and ICP4 fragmentsencoded by RS1.3.1 and RS1.3.2. OLPs were formulated in TiterMaxadjuvant (Alexis Biochemical) in a total volume of 100 μl per mousewhere adjuvant represented 1/3 of the subcutaneous dose. Mice wereimmunized on day 0, boosted on day 6 and spleens and blood werecollected on day 11. Single cell suspensions were prepared from spleensand erythrocytes were lysed. The splenocyte suspensions were thendivided into halves. The first half was separated into antigenpresenting cells, CD4⁺ and CD8⁺ T cells; 200,000 T cells were seeded perwell of IFN-gamma ELISPOT plate and stimulated with 100,000 APCs and OLPpool corresponding to immunization, irrelevant peptide, positive andnegative control. Cells were incubated in plates overnight after whichthe plates were developed and spots per well were counted. The secondhalf of each splenocyte suspension was run as unseparated splenocytes(400,000/well), pulsed with peptides, and assayed as described above.Results are shown in FIGS. 2A and B as magnitude of response perimmunization group.

G. [Protocol G] Vaccination with at Least Two Antigens

Example 1 Immunogenicity of gD2ΔTMR and ICP4 or ICP4.2 in C57BL/6 Mice

Purified protein was mixed with adjuvant and immunized into naïve miceto evaluate the ability to make CD4⁺ and CD8⁺ T cell responses to theprotein antigens. Briefly, antigen alone (gD2ΔTMR (5 μg)) orcombinations of antigens (gD2ΔTMR and ICP4.2 (10 μg)) were mixed withadjuvant (12 μg dose of an ISCOM matrix with a 82:18 mixture of Quillajasaponin fractions A and C) and administered subcutaneously to mice,twice, 9 days apart. Seven days after the second immunization, mice wereeuthanized and spleens were harvested for ex vivo IFNγ ELISPOT assays.CD4⁺ and CD8⁺ T cells were sorted out of the splenocyte population usingantibody-coated magnetic beads and then co-cultured on IFNγ-specificantibody-coated membranes in 97-well plates with naïve splenocytes thatwere pulsed with specific or non-specific antigens (as described) andirradiated with an x-ray irradiator. After 18 hours of incubation,captured IFNγ was detected with a biotinylated secondary IFNγ-specificantibody and visualized with horseradish peroxidase and3-amino-9-ethylcarbazole substrate. Data are reported as the number ofIFN-γ spot forming units per 2×10⁵ T cells±standard deviation of threemice per group. FIG. 3 shows the number of IFN-γ spot forming units per2×10⁵ CD4⁺ or CD8⁺ T cells±standard deviation of three mice per group.As seen in FIGS. 3A and B, the number of IFN-γ spot forming units perCD4⁺ T cells or CD8⁺ T cells is increased in mice immunized with gD2ΔTMRantigen combined with ICP4.2 compared to gD2ΔTMR antigen alone.

Example 2 Combinations of gD2 and ICP4.2 Plus Adjuvant ImmunizationReduced Disease Symptoms and Mortality in Mice

The ability to trigger protective immunity after immunization with theICP4.2 protein in combination with gD2 plus adjuvant was evaluated in alethal HSV-2 challenge mouse model. Briefly, eight C57BL/6 mice pergroup were immunized with either gD2 (2 μg) or ICP4.2 (10 μg) plusadjuvant individually or with both antigens mixed together plusadjuvant. Formulations were administered subcutaneously in the scruff ofthe neck twice, 9 days apart. Estrus cycles were synchronized with depoprovera 5 days prior to virus infection, and animals were challengedintravaginally 7 days after the second immunization with 20 times theLD₅₀ of HSV-2 strain 333. Disease symptoms were scored post-infection,and survival monitored. Disease severity scores were as follows: 0=nosymptoms, 1=redness, 2=redness and swelling, 3=herpetic lesions,4=severe ulceration or unilateral paralysis, and 5=bilateral paralysisor death.

TABLE 14 Effect of HSV-2 proteins gD2 and ICP4.2 on disease symptoms,viral replication and mortality Mean Reduc- Reduc- Antigen disease tionin tion in % (+adjuvant) score disease P virus Survival N = 8 Day 7score value** titer Day 11 PBS 3.5 ± 0.3 — — —  0% gD2* (2 ug) 2.5 ± 0.229% 0.016 0% 25% ICP4.2 (10 ug) 1.7 ± 0.4 51% 0.005 0% 13% gD2 (2 ug) +1.3 ± 0.3 63% 0.0004 20%  50% ICP4.2 (10 ug) *EC; **Student's t-test

Example 3 Combinations of gD2ΔTMR and ICP4.2 Plus Adjuvant ImmunizationReduced Disease Symptoms and Mortality in Mice

Mice immunized with a combination of gD2ΔTMR and ICP4.2 antigens showeda lower mean disease score at ten days after virus challenge compared toanimals receiving the individual antigen with adjuvant.

TABLE 15 Effect of HSV-2 proteins gD2ΔTMR and ICP4.2 on disease symptomsand survival rate in mice Mean Disease % Score % Reduc- P SurvivalGroups Day 10 tion value* Day 12 Adjuvant only 4.81 — —  0% gD2ΔTMR +adjuvant 1.44 70 0.023 75% gD2ΔTMR + ICP4.2 + 0.75 84 0.020 88% adjuvant

Example 4 Combination of gD2 and ICP4.2 Plus Adjuvant ImmunizationReduces Severity of Recurrent Lesions when Administered Therapeuticallyto HSV-2 Infected Guinea Pigs

The ability to affect HSV-2 reactivation in infected guinea pigs aftertherapeutic immunization with antigens plus adjuvant was evaluated.Briefly, guinea pigs were infected intravaginally with 5×10⁵ pfu ofHSV-2 strain MS, monitored for primary disease for 14 days, and thenrandomized into immunization groups (N=15). Animals were immunized threetimes subcutaneously on day 14, 21, and 35 post-infection with antigen(15 μg) plus adjuvant (50 μg) or adjuvant alone, or vehicle control andscored daily for local disease severity. The scoring system was asfollows: 0=no symptoms, 1=redness, 2=single lesions, 3=large or fusedlesions, 4=severe ulceration or unilateral paralysis, and 5=bilateralparalysis or death. Table 16 shows the data as the mean recurrent lesionscore for each week after the guinea pigs recovered from their acutedisease. The guinea pigs treated with a combination of gD2 and ICP4.2antigens showed a reduction in the mean lesion score at 7 (day 42) and14 (day 49) days after their last immunization compared to animalsreceiving the individual antigens with adjuvant.

TABLE 16 Effect of HSV-2 proteins gD2 and ICP4.2 vaccine on recurrentgenital skin disease Mean Recurrent Lesion Score Post HSV-2 InfectionAntigen + Adjuvant Day 15-21 Day 22-28 Day 29-35 Day 36-42 Day 43-49 PBS2.00 ± 0.45 1.17 ± 0.35 1.50 ± 0.50 0.87 ± 0.28 1.33 ± 0.33 gD2 1.00 ±0.30 0.67 ± 0.24 0.80 ± 0.19 0.83 ± 0.26 0.77 ± 0.28 ICP4.2 1.97 ± 0.381.07 ± 0.29 1.03 ± 0.33 0.53 ± 0.16 0.83 ± 0.29 gD2 & ICP4.2 1.43 ± 0.320.80 ± 0.27 1.07 ± 0.33 0.43 ± 0.19 0.70 ± 0.27

Sequences SEQ ID NO: 1 = ICP4, full-lengthSAEQRKKKKTTTTTQGRGAEVAMADEDGGRLRAAAETTGGPGSPDPADGPPPTPNPDRRPAARPGFGWHGGPEENEDEADDAAADADADEAAPASGEAVDEPAADGVVSPRQLALLASMVDEAVRTIPSPPPERDGAQEEAARS PSPPRTPSMRADYGEENDDDDDDDDDDDRDAGRWVRGPETTSAVRGAYPDPMASLSPRPPAPRRHHHHHHHRRRRAPRRRSAASDSSKSGSSSSASSASSSASSSSSASASSSDDDDDDDAARAPASAADHAAGGTLGADDEEAGVPARAPGAAPRPSPPRAEPAPARTPAATAGRLERRRARAAVAGRDATGRFTAGRPRRVELDADAASGAFYARYRDGYVSGEPWPGAGPPPPGRVLYGGLGDSRPGLWGAPEAEEARARFEASGAPAPVWAPELGDAAQQYALITRLLYTPDAEAMGWLQNPRVAPGDVALDQACFRISGAARNSSSFISGSVARAVPHLGYAMAAGRFGWGLAHVAAAVAMSRRYDRAQKGFLLTSLRRAYAPLLARENAALTGARTPDDGGDANRHDGDDARGKPAAAAAPLPSAAASPADERAVPAGYGAAGVLAALGRLSAAPASAPAGADDDDDDDGAGGGGGGRRAEAGRVAVECLAACRGILEALAEGFDGDLAAVPGLAGARPAAPPRPGPAGAAAPPHADAPRLRAWLRELRFVRDALVLMRLRGDLRVAGGSEAAVAAVRAVSLVAGALGPALPRSPRLLSSAAAAAADLLFQNQSLRPLLADTVAAADSLAAPASAPREARKRKSPAPARAPPGGAPRPPKKSRADAPRPAAAPPAGAAPPAPPTPPPRPPRPAALTRRPAEGPDPQGGWRRQPPGPSHTPAPSAAALEAYCAPRAVAELTDHPLFPAPWRPALMFDPRALASLAARCAAPPPGGAPAAFGPLRASGPLRRAAAWMRQVPDPEDVRVVILYSPLPGEDLAAGRAGGGPPPEWSAERGGLSCLLAALGNRLCGPATAAWAGNWTGAPDVSALGAQGVLLLSTRDLAFAGAVEFLGLLAGACDRRLIVVNAVRAADWPADGPVVSRQHAYLACEVLPAVQCAVRWPAARDLRRTVLASGRVFGPGVFARVEAAHARLYPDAPPLRLCRGANVRYRVRTRFGPDTLVPMSPREYRRAVLPALDGRAAASGAGDAMAPGAPDFCEDEAHSHRACARWGLGAPLRPVYVALGRDAVRGGPAELRGPRREFCARALLEPDGDAPPLVLRDDADAGPPPQIRWASAAGRAGTVLAAAGGGVEVVGTAAGLATPPRREPVDMDAELEDDDDGLFGE SEQ ID NO: 2 = ICP4 internal fragmentMVLYGGLGDSRPGLWGAPEAEEARARFEASGAPAPVWAPELGDAAQQYALITRLLYTPDAEAMGWLQNPRVAPGDVALDQACFRISGAARNSSSFISGSVARAVPHLGYAMAAGRFGWGLAHVAAAVAMSRRYDRAQKGFLLTS LRRAYAPLLARENAALTGARTPDDGGDANRRDGDDARGKPAAAAAPLPSAAASPADERAVPAGYGAAGVLAALGRLS AAPASAPAGADDDDDDDDGAGGGGGGGGGGGGRRAEAGRVAVECLAACRGILEALAEGFDGDLAAVPGLAGARPAAPPRPGPAGAAAPPHADAPRLRAWLRELRFVRDALVLMRLRGDLRVAGGSEAAVAAVRAVSLVAGALGPALPRSPRLLS SAAAAAADLLFQNQSL SEQ ID NO: 3 = gL2 cytoplasmicMGFVCLFGLVVMGAWGAWGGSQATEYVLRSVIAKEVGDILRVPCMRTPADDVSWRYEAPSVIDYARIDGIFLRYHCPGLDTFLWDRHAQRAYLVNPFLFAAGFLEDLSHSVFPADTQETTTRRALYKEIRDALGSRKQAVSHAPVRAGCVNFDYSRTRRCVGRRDLRPANTTSTWEPPVSSDDEASSQSKPLATQPPVLALSNAPPRRVSPTRGRRRHTRLRRN SEQ ID NO: 4 = gD2 internal deletionNRWKYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPSIPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFTPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHHAPAAPSNPRRRAQMAPKRLRLPHIRDDDAPPSHQPLFYSEQ ID NO: 5 = predicted gD2 sequenceMGRLTSGVGTAALLVVAVGLRVVCAKYALADPSLKMADPNRFRGKNLPVLDQLTDPPGVKRVYHIQPSLEDPFQPPS IPITVYYAVLERACRSVLLHAPSEAPQIVRGASDEARKHTYNLTIAWYRMGDNCAIPITVMEYTECPYNKSLGVCPIRTQPRWSYYDSFSAVSEDNLGFLMHAPAFETAGTYLRLVKINDWTEITQFILEHRARASCKYALPLRIPPAACLTSKAYQQGVTVDSIGMLPRFTPENQRTVALYSLKIAGWHGPKPPYTSTLLPPELSDTTNATQPELVPEDPEDSALLEDPAGTVSSQIPPNWHIPSIQDVAPHHAPAAPSNPGLIIGALAGSTLAVLVIGGIAFWVRRRAQMAPKRLRLPHIRDDDAPPSHQPLFY SEQ ID NO: 6 = ICP34.5MSRRRGPRRRGPRRRPRPGAPAVPRPGAPAVPRPGALPTADSQMVPAYDSGTAVESAPAASSLLRRWLLVPQADDSDDADYAGNDDAEWANSPPSEGGGKAPEAPHAAPAAACPPPPPRKERGPQRPLPPHLALRLRTTTEYLARLSLRRRRPPASPPADAPRGKVCFSPRVQVRHLVAWETAARLARRGSWARERADRDRFRRRVAAAEAVIGPCLEPEARARARARARAHEDGGPAEEEEAAAAARGSSAAAGPGRRAV SEQ ID NO: 7 = ICP0MEPRPGTSSRADPGPERPPRQTPGTQPAAPHAWGMLNDMQWLASSDSEEETEVGISDDDLHRDSTSEAGSTDTEMFEAGLMDAATPPARPPAERQGSPTPADAQGSCGGGPVGEEEAEAGGGGDVCAVCTDEIAPPLRCQSFPCLHPFCIPCMKTWIPLRNTCPLCNTPVAYLIVGVTASGSFSTIPIVNDPRTRVEAEAAVRAGTAVDFIWTGNPRTAPRSLSLGGHTVRALSPTPPWPGTDDEDDDLADVDYVPPAPRRAPRRGGGGAGATRGTSQPAATRPAPPGAPRSSSSGGAPLRAGVGSGSGGGPAVAAVVPRVASLPPAAGGGRAQARRVGEDAAAAEGRTPPARQPRAAQEPPIVISDSPPPSPRRPAGPGPLSFVSSSSAQVSSGPGGGGLPQSSGRAARPRAAVAPRVRSPPRAAAAPVVSASADAAGPAPPAVPVDAHRAPRSRMTQAQTDTQAQSLGRAGATDARGSGGPGAEGGPGVPRGTNTPGAAPHAAEGAAARPRKRRGSDSGPAASSSAS SSAAPRSPLAPQGVGAKRAAPRRAPDSDSGDRGHGPLAPASAGAAPPSASPSSQAAVAAASSSSASSSSASSSSASS SSASSSSASSSSASSSSASSSAGGAGGSVASASGAGERRETSLGPRAAAPRGPRKCARKTRHAEGGPEPGARDPAPGLTRYLPIAGVSSVVALAPYVNKTVTGDCLPVLDMETGHIGAYVVLVDQTGNVADLLRAAAPAWSRRTLLPEHARNCVRPPDYPTPPASEWNSLWMTPVGNMLFDQGTLVGALDFHGLRSRHPWSREQGAPAPAGDAPAGHGESEQ ID NO: 8 = ICP4 internal fragments (RS1.1, #1-400)msaeqrkkkktttttqgrgaevamadedggrlraaaettggpgspdpadgppptpnpdrrpaarpgfgwhggpeenedeaddaaadadadeaapasgeavdepaadgvvsprqlallasmvdeavrtipsppperdgaqeeaarspspprtpsmradygeendddddddddddrdagrwvrgpettsavrgaypdpmaslsprppaprrhhhhhhhrrrraprrrsaasdssksgssssassasssasssssasasssdddddddaarapasaadhaaggtlgaddeeagvparapgaaprpsppraepapartpaatagrlerrraraavagrdatgrftagrprrveldadaasgafyaryrdgyvsgepwpgagppppgrvlygglgdsrpglwgap SEQ ID NO: 9 =ICP4 internal framents (RS1.3.1, #750-1024)ssaaaaaadllfqnqslrplladtvaaadslaapasaprearkrkspaparappggaprppkksradaprpaaappagaappapptppprpprpaaltrrpaegpdpqggwrrqppgpshtpapsaaaleaycapravaeltdhplfpapwrpalmfdpralaslaarcaapppggapaafgplrasgplrraaawmrqvpdpedvrvvilysplpgedlaagragggpppewsaergglscllaalgnrlcgpataawagnwtgapdvsalgaq SEQ ID NO: 10 =ICP4 internal fragments (RS1.3.2, #1008-1319)wagnwtgapdvsalgaqgvlllstrdlafagaveflgllagacdrrlivvnavraadwpadgpvvsrqhaylacevlpavqcavrwpaardlrrtvlasgrvfgpgvfarveaaharlypdapplrlcrganvryrvrtrfgpdtlvpmspreyrravlpaldgraaasgagdamapgapdfcedeahshracarwglgaplrpvyvalgrdavrggpaelrgprrefcarallepdgdapplvlrddadagpppqirwasaagragtvlaaagggvevvgtaaglatpprrepvdmdaeleddddglfge SEQ ID NO: 11 =ICP4 internal fragments (RS1.3, #750-1319)SsaaaaaadllfqnqslrplladtvaaadslaapasaprearkrkspaparappggaprppkksradaprpaaappagaappapptppprpprpaaltrrpaegpdpqggwrrqppgpshtpapsaaaleaycapravaeltdhplfpapwrpalmfdpralaslaarcaapppggapaafgplrasgplrraaawmrqvpdpedvrvvilysplpgedlaagragggpppewsaergglscllaalgnrlcgpataawagnwtgapdvsalgaqgvlllstrdlafagaveflgllagacdrrlivvnavraadwpadgpvvsrqhaylacevlpavqcavrwpaardlrrtvlasgrvfgpgvfarveaaharlypdapplrlcrganvryrvrtrfgpdtlvpmspreyrravlpaldgraaasgagdamapgapdfcedeahshracarwglgaplrpvyvalgrdavrggpaelrgprrefcarallepdgdapplvlrddadagpppqirwasaagragtvlaaagggvevvgtaaglatpprrepvdmdaeleddddglfgeSEQ ID NO: 12 = ICP4 internal fragments (RS1.4, #340-883)tagrprrveldadaasgafyaryrdgyvsgepwpgagppppgrvlygglgdsrpglwgapeaeeararfeasgapapvwapelgdaaqqyalitrllytpdaeamgwlqnprvapgdvaldqacfrisgaarnsssfisgsvaravphlgyamaagrfgwglahvaaavamsrrydraqkgflltslrrayapllarenaaltgartpddggdanrhdgddargkpaaaaaplpsaaaspaderavpagygaagvlaalgrlsaapasapagadddddddgaggggggrraeagrvaveclaacrgilealaegfdgdlaavpglagarpaapprpgpagaaapphadaprlrawlrelrfvrdalvlmrlrgdlrvaggseaavaavravslvagalgpalprsprllssaaaaaadllfqnqslrplladtvaaadslaapasaprearkrkspaparappggaprppkksradaprpaaappagaappapptppprpprpaaltrrpaegpdpqggwrrqppgpshtpapsaaaleayca SEQ ID NO: 13 =ICP4 internal fragments (RS1.5, #775-1318)aaadslaapasaprearkrkspaparappggaprppkksradaprpaaappagaappapptppprpprpaaltrrpaegpdpqggwrrqppgpshtpapsaaaleaycapravaeltdhplfpapwrpalmfdpralaslaarcaapppggapaafgplrasgplrraaawmrqvpdpedvrvvilysplpgedlaagragggpppewsaergglscllaalgnrlcgpataawagnwtgapdvsalgaqgvlllstrdlafagaveflgllagacdrrlivvnavraadwpadgpvvsrqhaylacevlpavqcavrwpaardlrrtvlasgrvfgpgvfarveaaharlypdapplrlcrganvryrvrtrfgpdtlvpmspreyrravlpaldgraaasgagdamapgapdfcedeahshracarwglgaplrpvyvalgrdavrggpaelrgprrefcarallepdgdapplvlrddadagpppqirwasaagragtvlaaagggvevvgtaaglatpprrepvdmdaeleddddglfge SEQ ID NO: 14 =ICP4 internal fragments (RS1.6, #209-1318)hhhhhhhrrrraprrrsaasdssksgssssassasssasssssasasssdddddddaarapasaadhaaggtlgaddeeagvparapgaaprpsppraepapartpaatagrlerrraraavagrdatgrftagrprrveldadaasgafyaryrdgyvsgepwpgagppppgrvlygglgdsrpglwgapeaeeararfeasgapapvwapelgdaaqqyalitrllytpdaeamgwlqnprvapgdvaldqacfrisgaarnsssfisgsvaravphlgyamaagrfgwglahvaaavamsrrydraqkgflltslrrayapllarenaaltgartpddggdanrhdgddargkpaaaaaplpsaaaspaderavpagygaagvlaalgrlsaapasapagadddddddgaggggggrraeagrvaveclaacrgilealaegfdgdlaavpglagarpaapprpgpagaaapphadaprlrawlrelrfvrdalvlmrlrgdlrvaggseaavaavravslvagalgpalprsprllssaaaaaadllfqnqslrplladtvaaadslaapasaprearkrkspaparappggaprppkksradaprpaaappagaappapptppprpprpaaltrrpaegpdpqggwrrqppgpshtpapsaaaleaycapravaeltdhplfpapwrpalmfdpralaslaarcaapppggapaafgplrasgplrraaawmrqvpdpedvrvvilysplpgedlaagragggpppewsaergglscllaalgnrlcgpataawagnwtgapdvsalgaqgvlllstrdlafagaveflgllagacdrrlivvnavraadwpadgpvvsrqhaylacevlpavqcavrwpaardlrrtvlasgrvfgpgvfarveaaharlypdapplrlcrganvryrvrtrfgpdtlvpmspreyrravlpaldgraaasgagdamapgapdfcedeahshracarwglgaplrpvyvalgrdavrggpaelrgprrefcarallepdgdapplvlrddadagpppqirwasaagragtvlaaagggvevvgtaaglatpprrepvdmdaeleddddglfge SEQ ID NO: 15 =ICP4 internal fragments (RS1.7, deletion of 391-544)msaeqrkkkktttttqgrgaevamadedggrlraaaettggpgspdpadgppptpnpdrrpaarpgfgwhggpeenedeaddaaadadadeaapasgeavdepaadgvvsprqlallasmvdeavrtipsppperdgaqeeaarspspprtpsmradygeendddddddddddrdagrwvrgpettsavrgaypdpmaslsprppaprrhhhhhhhrrrraprrrsaasdssksgssssassasssasssssasasssdddddddaarapasaadhaaggtlgaddeeagvparapgaaprpsppraepapartpaatagrlerrraraavagrdatgrftagrprrveldadaasgafyaryrdgyvsgepwpgagppppgrvlygglgartpddggdanrhdgddargkpaaaaaplpsaaaspaderavpagygaagvlaalgrlsaapasapagadddddddgaggggggrraeagrvaveclaacrgilealaegfdgdlaavpglagarpaapprpgpagaaapphadaprlrawlrelrfvrdalvlmrlrgdlrvaggseaavaavravslvagalgpalprsprllssaaaaaadllfqnqslrplladtvaaadslaapasaprearkrkspaparappggaprppkksradaprpaaappagaappapptppprpprpaaltrrpaegpdpqggwrrqppgpshtpapsaaaleaycapravaeltdhplfpapwrpalmfdpralaslaarcaapppggapaafgplrasgplrraaawmrqvpdpedvrvvilysplpgedlaagragggpppewsaergglscllaalgnrlcgpataawagnwtgapdvsalgaqgvlllstrdlafagaveflgllagacdrrlivvnavraadwpadgpvvsrqhaylacevlpavqcavrwpaardlrrtvlasgrvfgpgvfarveaaharlypdapplrlcrganvryrvrtrfgpdtivpmspreyrravlpaldgraaasgagdamapgapdfcedeahshracarwglgaplrpvyvalgrdavrggpaelrgprrefcarallepdgdapplvlrddadagpppqirwasaagragtvlaaagggvevvgtaaglatpprrepvdmdaeleddddglfge SEQ ID NO: 16 =ICP4 internal fragments (RS1.8, deletion of 786-864)msaeqrkkkktttttqgrgaevamadedggrlraaaettggpgspdpadgppptpnpdrrpaarpgfgwhggpeenedeaddaaadadadeaapasgeavdepaadgvvsprqlallasmvdeavrtipsppperdgageeaarspspprtpsmradygeendddddddddddrdagrwvrgpettsavrgaypdpmaslsprppaprrhhhhhhhrrrraprrrsaasdssksgssssassasssasssssasasssdddddddaarapasaadhaaggtlgaddeeagvparapgaaprpsppraepapartpaatagrlerrraraavagrdatgrftagrprrveldadaasgafyaryrdgyvsgepwpgagppppgrvlygglgdsrpglwgapeaeeararfeasgapapvwapelgdaaqqyalitrllytpdaeamgwlqnprvapgdvaldqacfrisgaarnsssfisgsvaravphlgyamaagrfgwglahvaaavamsrrydraqkgflltslrrayapllarenaaltgartpddggdanrhdgddargkpaaaaaplpsaaaspaderavpagygaagvlaalgrlsaapasapagadddddddgaggggggrraeagrvaveclaacrgilealaegfdgdlaavpglagarpaapprpgpagaaapphadaprlrawlrelrfvrdalvlmrlrgdlrvaggseaavaavravslvagalgpalprsprllssaaaaaadllfqnqslrplladtvaaadslaapastpapsaaaleaycapravaeltdhplfpapwrpalmfdpralaslaarcaapppggapaafgplrasgplrraaawmrqvpdpedvrvvilysplpgedlaagragggpppewsaergglscllaalgnrlcgpataawagnwtgapdvsalgaqgvlllstrdlafagaveflgllagacdrrlivvnavraadwpadgpvvsrqhaylacevlpavqcavrwpaardlrrtvlasgrvfgpgvfarveaaharlypdapplrlcrganvryrvrtrfgpdtlvpmspreyrravlpaldgraaasgagdamapgapdfcedeahshracarwglgaplrpvyvalgrdavrggpaelrgprrefcarallepdgdapplvlrddadagpppqirwasaagragtvlaaagggvevvgtaaglatpprrepvdmdaeleddddglfge SEQ ID NO: 17 =predicted sequence for uracil DNA glycosylase (encoded by UL2)MFSASTTPEQPLGLSGDATPPLPTSVPLDWAAFRRAFLIDDAWRPLLEPELANPLTARLLAEYDRRCQTEEVLPPREDVFSWTRYCTPDDVRVVIIGQDPYHHPGQAHGLAFSVRADVPVPPSLRNVLAAVKNCYPDARMSGRGCLEKWARDGVLLLNTTLTVKRGAAASHSKLGWDRFVGGVVQRLAARRPGLVFMLWGAHAQNAIRPDPRQHYVLKFSHPSPLSKVPFGTCQHFLAANRYLETRDIMPIDWSV SEQ ID NO: 18 =predicted sequence for tegument protein encoded by UL11MGLAFSGARPCCCRHNVITTDGGEVVSLTAHEFDVVDIESEEEGNFYVPPDVRVVTRAPGPQYRRASDPPSRHTRRRDPDVARPPATLTPPLSDSE SEQ ID NO: 19 = gL2 secretedNRWGFVCLFGLVVMGAWGAWGGSQATEYVLRSVIAKEVGDILRVPCMRTPADDVSWRYEAPSVIDYARIDGIFLRYHCPGLDTFLWDRHAQRAYLVNPFLFAAGFLEDLSHSVFPADTQETTTRRALYKEIRDALGSRKQAVSHAPVRAGCVNFDYSRTRRCVGRRDLRPANTTSTWEPPVSSDDEASSQSKPLATQPPVLALSNAPPRRVSPTRGRRRHTRLRRN SEQ ID NO: 20 =predicted sequence for VP5 encoded by UL19DYDIPTTENLYFQGMAAPARDPPGYRYAAAMVPTGSILSTIEVASHRRLFDFFARVRSDENSLYDVEFDALLGSYCNTLSLVRFLELGLSVACVCTKFPELAYMNEGRVQFEVHQPLIARDGPHPVEQPVHNYMTKVIDRRALNAAFSLATEAIALLTGEALDGTGISLHRQLRAIQQLARNVQAVLGAFERGTADQMLHVLLEKAPPLALLLPMQRYLDNGRLATRVARATLVAELKRSFCDTSFELGKAGHRREAIEAWLVDLTTATQPSVAVPRLTHADTRGRPVDGVLVTTAAIKQRLLQSFLKVEDTEADVPVTYGEMVLNGANLVTALVMGKAVRSLDDVGRHLLEMQEEQLEANRETLDELESAPQTTRVRADLVAIGDRLVFLEALEKRIYAATNVPYPLVGAMDLTFVLPLGLFNPAMERFAAHAGDLVPAPGHPEPRAFPPRQLFFWGKDHQVLRLSMENAVGTVCHPSLMNIDAAVGGVNHDPVEAANPYGAYVAAPAGPGADMQQRFLNAWRQRLAHGRVRWVAECQMTAEQFMQPDNANLALELHPAFDFFAGVADVELPGGEVPPAGPGAIQATWRVVNGNLPLALCPVAFRDARGLELGVGRHAMAPATIAAVRGAFEDRSYPAVFYLLQAAIHGSEHVFCALARLVTQCITSYWNNTRCAAFVNDYS LVSYIVTYLGGDLPEECMAVYRDLVAHVEALAQLVDDFTLPGPELGGQAQAELNHLMRDPALLPPLVWDCDGLMRHAALDRHRDCRIDAGEHEPVYAAACNVATADFNRNDGRLLHNTQARAADAADDRPHRPADWTVHHKIYYYVLVPAFSRGRCCTAGVRFDRVYATLQNMVVPEIAPGEECPSDPVTDPAHPLHPANLVANTVNAMFHNGRVVVDGPAMLTLQVLAHNMAERTTALLCSAAPDAGANTASTANMRIFDGALHAGVLLMAPQHLDHTIQNGEYFYVLPVHALFAGADHVANAPNFPPALRDLARHVPLVPPALGANYFSSIRQPVVQHARESAAGENALTYALMAGYFKMSPVALYHQLKTGLHPGFGFTVVRQDRFVTENVLFSERASEAYFLGQLQVARHETGGGVSFTLTQPRGNVDLGVGYTAVAATATVRNPVTDMGNLPQNFYLGRGAPPLLDNAAAVYLRNAVVAGNRLGPAQPLPVFGCAQVPRRAGMDHGQDAVCEFIATPVATDINYFRRPCNPRGRAAGGVYAGDKEGDVIALMYDHGQSDPARPFAATANPWASQRFSYGDLLYNGAYHLNGASPVLSPCFKFFTAADITAKHRCLERLIVETGSAVSTATAASDVQFKRPPGCRELVEDPCGLFQEAYPITCASDPALLRSARDGEAHARETHFTQYLIYDASPLKGLSL SEQ ID NO: 21 = VP5 encoded by UL19ΔTEVMAAPARDPPGYRYAAAMVPTGSILSTIEVASHRRLFDFFARVRSDENSLYDVEFDALLGSYCNTLSLVRFLELGLSVACVCTKFPELAYMNEGRVQFEVHQPLIARDGPHPVEQPVHNYMTKVIDRRALNAAFSLATEAIALLTGEALDGTGISLHRQLRAIQQLARNVQAVLGAFERGTADQMLHVLLEKAPPLALLLPMQRYLDNGRLATRVARATLVAELKRSFCDTSFFLGKAGHRREAIEAWLVDLTTATQPSVAVPRLTHADTRGRPVDGVLVTTAAIKQRLLQSFLKVEDTEADVPVTYGEMVLNGANLVTALVMGKAVRSLDDVGRHLLEMQEEQLEANRETLDELESAPQTTRVRADLVAIGDRLVFLEALEKRIYAATNVPYPLVGAMDLTFVLPLGLFNPAMERFAAHAGDLVPAPGHPEPRAFPPRQLFFWGKDHQVLRLSMENAVGTVCHPSLMNIDAAVGGVNHDPVEAANPYGAYVAAPAGPGADMQQRFLNAWRQRLAHGRVRWVAECQMTAEQFMQPDNANLALELHPAFDFFAGVADVELPGGEVPPAGPGAIQATWRVVNGNLPLALCPVAFRDARGLELGVGRHAMAPATIAAVRGAFEDRSYPAVFYLLQAAIHGSEHVFCALARLVTQCITSYWNNTRCAAFVNDYSLVSYIVTYLGGDLPEECMAVYRDLVAHVEALAQLVDDFTLPGPELGGQAQAELNHLMRDPALLPPLVWDCDGLMRHAALDRHRDCRIDAGEHEPVYAAACNVATADFNRNDGRLLHNTQARAADAADDRPHRPADWTVHHKIYYYVLVPAFSRGRCCTAGVRFDRVYATLQNMVVPEIAPGEECPSDPVTDPAHPLHPANLVANTVNAMFHNGRVVVDGPAMLTLQVLAHNMAERTTALLCSAAPDAGANTASTANMRIFDGALHAGVLLMAPQHLDHTIQNGEYFYVLPVHALFAGADHVANAPNFPPALRDLARHVPLVPPALGANYFSSIRQPVVQHARESAAGENALTYALMAGYFKMSPVALYHQLKTGLHPGFGFTVVRQDRFVTENVLFSERASEAYFLGQLQVARHETGGGVSFTLTQPRGNVDLGVGYTAVAATATVRNPVTDMGNLPQNFYLGRGAPPLLDNAAAVYLRNAVVAGNRLGPAQPLPVFGCAQVPRRAGMDHGQDAVCEFIATPVATDINYFRRPCNPRGRAAGGVYAGDKEGDVIALMYDHGQSDPARPFAATANPWASQRFSYGDLLYNGAYHLNGASPVLSPCFKFFTAADITAKHRCLERLIVETGSAVSTATAASDVQFKRPPGCRELVEDPCGLFQEAYPITCASDPALLRSARDGEAHARETHFTQYLIYDASPLKGLSLSEQ ID NO: 22 = predicted sequence for ICP1/2 encoded by UL36MIPAALPHPTMKRQGDRDIVVTGVRNQFATDLEPGGSVSCMRSSLSFLSLLFDVGPRDVLSAEAIEGCLVEGGEWTRAAAGSGPPRMCSIIELPNFLEYPAARGGLRCVFSRVYGEVGFFGEPTAGLLETQCPAHTFFAGPWAMRPLSYTLLTIGPLGMGLYRDGDTAYLFDPHGLPAGTPAFIAKVRAGDVYPYLTYYAHDRPKVRWAGAMVFFVPSGPGAVAPADLTAAALHLYGASETYLQDEPFVERRVAITHPLRGEIGGLGALFVGVVPRGDGEGSGPVVPALPAPTHVQTPGADRPPEAPRGASGPPDTPQAGHPNRPPDDVWAAALEGTPPAKPSAPDAAASGPPHAAPPPQTPAGDAAEEAEDLRVLEVGAVPVGRHRARYSTGLPKRRRPTWTPPSSVEDLTSGERPAPKAPPAKAKKKSAPKKKAPVAAEVPASSPTPIAATVPPAPDTPPQSGQGGGDDGPASPSSPSVLETLGARRPPEPPGADLAQLFEVHPNVAATAVRLAARDAALAREVAACSQLTINALRS PYPAHPGLLELCVIFFFERVLAFLIENGARTHTQAGVAGPAAALLDFTLRMLPRKTAVGDFLASTRMSLADVAAHRPLIQHVLDENSQIGRLALAKLVLVARDVIRETDAFYGDLADLDLQLRAAPPANLYARLGEWLLERSRAHPNTLFAPATPTHPEPLLHRIQALAQFARGEEMRVEAEAREMREALDALARGVDSVSQRAGPLTVMPVPAAPGAGGRAPCPPALGPEAIQARLEDVRIQARRAIESAVKEYFHRGAVYSAKALQASDSHDCRFHVASAAVVPMVQLLESLPAFDQHTRDVAQRAALPPPPPLATSPQAILLRDLLQRGQPLDAPEDLAAWLSVLTDAATQGLIERKPLEELARSIHGINDQQARRSSGLAELQRFDALDAALAQQLDSDAAFVPATGPAPYVDGGGLSPEATRMAEDALRQARAMEAAKMTAELAPEARSRLRERAHALEAMLNDARERAKVAHDAREKFLHKLQGVLRPLPDFVGLKACPAVLATLRASLPAGWTDLADAVRGPPPEVTAALRADLWGLLGQYREALEHPTPDTATALAGLHPAFVVVLKTLFADAPETPVLVQFFSDHAPTIAKAVSNAINAGSAAVATASPAATVDAAVRAHGALADAVSALGAAARDPASPLSFLAVLADSAAGYVKATRLALEARGAIDELTTLGSAAADLVVQARRACAQPEGDHAALIDAAARATTAARESLAGHEAGFGGLLHAEGTAGDHSPSGRALQELGKVIGATRRRADELEAAVADLTAKMAAQRARGSSERWAAGVEAALDRVENRAEFDVVELRRLQALAGTHGYNPRDFRKRAEQALAANAEAVTLALDTAFAFNPYTPENQRHPMLPPLAAIHRLGWSAAFHAAAETYADMFRVDAEPLARLLRIAEGLLEMAQAGDGFIDYHEAVGRLADDMTSVPGLRRYVPFFQHGYADYVELRDRLDAIRADVHRALGGVPLDLAAAAEQISAARNDPEATAELVRTGVTLPCPSEDALVACAAALERVDQSPVKNTAYAEYVAFVTRQDTAETKDAVVRAKQQRAEATERVMAGLREALAARERRAQIEAEGLANLKTMLKVVAVPATVAKTLDQARSVAEIADQVEVLLDQTEKTRELDVPAVIWLEHAQRTFETHPLSAARGDGPGPLARHAGRLGALFDTRRRVDALRRSLEEAEAEWDEVWGRFGRVRGGAWKSPEGFRAMHEQLRALQDTTNTVSGLRAQPAYERLSARYQGVLGAKGAERAEAVEELGARVTKHTALCARLRDEVVRRVPWEMNFDALGGLLAEFDAAAADLAPWAVEEFRGARELIQYRMGLYSAYARAGGQTGAGAESAPAPLLVDLRALDARARASSSPEGHEVDPQLLRRRGEAYLRAGGDPGPLVLREAVSALDLPFATSFLAPDGTPLQYALCFPAVTDKLGALLMRPEAACVRPPLPTDVLESAPTVTAMYVLTVVNRLQLALSDAQAANFQLFGRFVRHRQATWGASMDAAAELYVALVATTLTREFGCRWAQLGWASGAAAPRPPPGPRGSQRHCVAFNENDVLVALVAGVPEHIYNFWRLDLVRQHEYMHLTLERAFEDAAESMLFVQRLTPHPDARIRVLPTFLDGGPPTRGLLFGTRLADWRRGKLSETDPLAPWRSALELGTQRRDVPALGKLSPAQALAAVSVLGRMCLPSAALAALWTCMFPDDYTEYDSFDALLAARLESGQTLGPAGGREASLPEAPHALYRPTGQHVAVLAAATHRTPAARVTAMDLVLAAVLLGAPVVVALRNTTAFSRESELELCLTLFDSRPGGPDAALRDVVSSDIETWAVGLLHTDLNPIENACLAAQLPRLSALIAERPLADGPPCLVLVDISMTPVAVLWEAPEPPGPPDVRFVGSEATEELPFVATAGDVLAASAADADPFFARAILGRPFDASLLTGELFPGHPVYQRPLADEAGPSAPTAARDPRDLAGGDGGSGPEDPAAPPARQADPGVLAPTLLTDATTGEPVPPRMWAWIHGLEELASDDAGGPTPNPAPALLPPPATDQSVPTSQYAPRPIGPAATARETRPSVPPQQNTGRVPVAPRDDPRPSPPTPSPPADAALPPPAFSGSAAAFSAAVPRVRRSRRTRAKSRAPRASAPPEGWRPPALPAPVAPVAASARPPDQPPTPESAPPAWVSALPLPPGPASARGAFPAPTLAPIPPPPAEGAVVPGGDRRRGRRQTTAGPSPTPPRGPAAGPPRRLTRPAVASLSASLNSLPSPRDPADHAAAVSAAAAAVPPSPGLAPPTSAVQTSPPPLAPGPVAPSEPLCGWVVPGGPVARRPPPQSPATKPAARTRIRARSVPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPPVTRTLTPQSRDSVPTPESPTHTNTHLPVSAVTSWASSLALHVDSAPPPASLLQTLHISSDDEHSDADSLRFSDSDDTEALDPLPPEPHLPPADEPPGPLAADHLQSPHSQFGPLPVQANAVLSRRYVRSTGRS ALAVLIRACRRIQQQLQRTRRALFQRSNAVLTSLHHVRMLLG SEQ ID NO: 23 =ICP1/2 internal fragments encoded by UL36.3.4.1AAQRARGSSERWAAGVEAALDRVENRAEFDVVELRRLQALAGTHGYNPRDFRKRAEQALAANAEAVTLALDTAFAFNPYTPENQRHPMLPPLAAIHRLGWSAAFHAAAETYADMFRVDAEPLARLLRIAEGLLEMAQAGDGFIDYHEAVGRLADDMTSVPGLRRYVPFFQHGYADYVELRDRLDAIRADVHRALGGVPLDLAAAAEQISAARNDPEATAELVRTGVTLPCPSEDALVACAAALERVDQSPVKNTAYAEYVAFVTRQDTAETKDAVVRAKQQRAEATERVMAGLREALAARERRAQIEAEGLANLKTMLKVVAVPATVAKTLDQARSVAEIADQVEVLLDQTEKTRELDVPAVIWLEHAQRTFETHPLSAARGDGPGPLARHAGRLGALFDTRRRVDALRRSLEEAEAEWDEVWGRFGRVRGGAWKSPEGFRAMHEQLRALQDTTNTVSGLRAQPAYERLSARYQGVLGAKGAERAEAVEELGARVTKHTALCARLRDEVVRRVPWEMNFDALGGLLAEFDAAAADLAPWAVEEFRGARELIQYRMGLYSAYARAGGQTGAGAESAPAPLLVDLRALDARARASSSPEGHEVDPQLLRRRGEAYLRAGGDPGPLVLREAVSALDLPFATSFLAPDGTPLQYALCFPAVTDKLGALLMRPEAACVRPPLPTDVLESAPTVTAMYVLTVVNRLQLALSDAQAANFQLFGRFVRHRQATWGASMDAAAELYVALVATTLTREFGCRWAQLGWAS GAAAPRPPPGPRGSQRHCVAFNENDVLVALVAGVPEHIYNFWRLDLVRQHEYMHLTLERAFEDAAESMLFVQRLTPHPDARTRVLPTFLDGGPPTRGLLFGTRLADWRRGKLSETDPLAPWRSALELGTQRRDVPALGKLSPAQALAAVSVLGRMCLPSAALAALWTCMFPDDYTEYDSFDALLAARLESGQTLGPAGGREASL SEQ ID NO: 24 =ICP1/2 internal fragments encoded by UL36.4.2.5EYDSFDALLAARLESGQTLGPAGGREASLPEAPHALYRPTGQHVAVLAAATHRTPAARVTAMDLVLAAVLLGAPVVVALRNTTAFSRESELELCLTLFDSRPGGPDAALRDVVSSDIETWAVGLLHTDLNPIENACLAAQLPRLSALIAERPLADGPPCLVLVDISMTPVAVLWEAPEPPGPPDVRFVGSEATEELPFVATAGDVLAASAADADPFFARAILGRPFDASLLTGELFPGHPVYQRPLADEAGPSAPTAARDPRDLAGGDGGSGPEDPAAPPARQADPGVLAPTLLTDATTGEPVPPRMWAWIHGLEELASDDAGGPTPNPAPALLPPPATDQSVPTSQYAPRPIGPAATARETRPSVPPQQNTGRVPVAPRDDPRPSPPTPSPPADAALPPPAFSGSAAAFSAAVPRVRRSRRTRAKSRAPRASAPPEGWRPPALPAPVAPVAASARPPDQPPTPESAPPAWVSALPLPPGPASARGAFPAPTLAPIPPPPAEGAVVPGGDRRRGRRQTTAGPSPTPPRGPAAGPPRRLTRPAVASLSASLNSLPSPRDPADHAAAVSAAAAAVPPSPGLAPPTSAVQTSPPPLAPGPVAPSEPLCGWVVPGGPVARRPPPQSPATKPAARTRIRARSVPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPQPPLPPVTRTLTPQSRDSVPTPESPTHTNTHLPVSAVTSWASSLALHVDSAPPPASLLQTLHISSDDEHSDADSLRFSDSDDTEALDPLPPEPHLPPADEPPGPLAADHLQSPHSQFGPLPVQANAVLSRRYVRSTGRSALAVLIRACRRIQQQLQRTRRALFQRSNAVLTSLHHVRMLLG SEQ ID NO: 25 =predicted sequence for reductase encoded by UL40MDPAVSPASTDPLDTHASGAGAAPIPVCPTPERYFYTSQCPDINHLRSLSILNRWLETELVFVGDEEDVSKLSEGELGFYRFLFAFLSAADDLVTENLGGLSGLFEQKDILHYYVEQECIEVVHSRVYNIIQLVLFHNNDQARRAYVARTINHPAIRVKVDWLEARVRECDSIPEKFILMILIEGVFFAASFAAIAYLRTNNLLRVTCQSNDLISRDEAVHTTASCYIYNNYLGGHAKPEAARVYRLFREAVDIEIGFIRSQAPTDSSILSPGALAAIENYVRFSADRLLGLIHMQPLYSAPAPDASFPLSLMSTDKHTNFFECRSTSYAGAVVNDL SEQ ID NO: 26 =ICP47 encoded by U512MSWALKTTDMFLDSSRCTHRTYGDVCAEIHKREREDREAARTAVTDPELPLLCPPDVRSDPASRNPTQQTRGCARSNERQDRVLAP SEQ ID NO: 27 = gM2 encoded by UL10MGRRAPRGSPEAAPGADVAPGARAAWWVWCVQVATFIVSAICVVGLLVLASVFRDRFPCLYAPATSYAKANATVEVRGGVAVPLRLDTQSLLATYAITSTLLLAAAVYAAVGAVTSRYERALDAARRLAAARMAMPHATLIAGNVCAWLLQITVLLLAHRISQLAHLIYVLHFACLVYLAAHFCTRGVLSGTYLRQVHGLIDPAPTHHRIVGPVRAVMTNALLLGTLLCTAAAAVSLNTIAALNFNFSAPSMLICLTTLFALLVVSLLLVVEGVLCHYVRVLVGPHLGAIAATGIVGLACEHYHTGGYYVVEQQWPGAQTGVRVALALVAAFALAMAVLRCTRAYLYHRRHHTKFFVRMRDTRHRAHSALRRVRSSMRGSRRGGPPGDPGYAETPYASVSHHAEIDRYGDSDGDPIYDEVAPDHEAELYARVQRPGPVPDAEPIYDTVEGYAPRSAGEPVYS TVRRW SEQ ID NO: 28 =predicted sequence for cleavage/package protein encoded by UL15MFGQQLASDVQQYLERLEKQRQQKVGVDEASAGLTLGGDALRVPFLDFATATPKRHQTVVPGVGTLHDCCEHSPLFS AVARRLLFNSLVPAQLRGRDFGGDHTAKLEFLAPELVRAVARLRFRECAPEDAVPQRNAYYSVLNTFQALHRSEAFRQLVHFVRDFAQLLKTSFRASSLAETTGPPKKRAKVDVATHGQTYGTLELFQKMILMHATYFLAAVLLGDHAEQVNTFLRLVFEIPLFSDTAVRHFRQRATVFLVPRRHGKTWFLVPLIALSLASFRGIKIGYTAHIRKATEPVFDEIDACLRGWFGSSRVDHVKGETISFSFPDGSRSTIVFASSHNTNGIRGQDFNLLFVDEANFIRPDAVQTIMGFLNQANCKIIFVSS TNTGKASTSFLYNLRGAADELLNVVTYICDDHMPRVVTHTNATACSCYILNKPVFITMDGAVRRTADLFLPDSFMQEIIGGQARETGDDRPVLTKSAGERFLLYRPSTTTNSGLMAPELYVYVDPAFTANTRASGTGIAVVGRYRDDFIIFALEHFFLRALTGSAPADIARCVVHSLAQVLALHPGAFRSVRVAVEGNSSQDSAVAIATHVHTEMHRILASAGANGPGPELLFYHCEPPGGAVLYPFFLLNKQKTPAFEYFIKKFNSGGVMASQELVSVTVRLQTDPVEYLSEQLNNLIETVSPNTDVRMYSGKRNGAADDLMVAVIMAIYLAAPTGIPPAFFPITRTS  SEQ ID NO: 29 =predicted sequence for ICP35 encoded by UL26.5MNPVSASGAPAPPPPGDGSYLWIPASHYNQLVTGQSAPRHPPLTACGLPAAGTVAYGHPGAGPSPHYPPPPAHPYPGMLFAGPSPLEAQIAALVGAIAADRQAGGLPAAAGDHGIRGSAKRRRHEVEQPEYDCGRDEPDRDFPYYPGEARPEPRPVDSRRAARQASGPHETITALVGAVTSLQQELAHMRARTHAPYGPYPPVGPYHHPHADTETPAQPPRYPAKAVYLPPPHIAPPGPPLSGAVPPPSYPPVAVTPGPAPPLHQPSPAHAHPPPPPPGPTPPPAASLPQPEAPGAEAGALVNASSAAHVNVDTARAADLFVSQMMGSR SEQ ID NO: 30 =predicted sequence for polymerase encoded by UL30MFCAAGGPASPGGKPAARAASGFFAPHNPRGATQTAPPPCRRQNFYNPHLAQTGTQPKALGPAQRHTYYSECDEFRFIAPRSLDEDAPAEQRTGVHDGRLRRAPKVYCGGDERDVLRVGPEGFWPRRLRLWGGADHAPEGFDPTVTVFHVYDILEHVEHAYSMRAAQLHERFMDAITPAGTVITLLGLTPEGHRVAVHVYGTRQYFYMNKAEVDRHLQCRAPRDLCERLAAALRESPGASFRGISADHFEAEVVERADVYYYETRPTLYYRVFVRSGRALAYLCDNFCPAIRKYEGGVDATTRFILDNPGFVTFGWYRLKPGRGNAPAQPRPPTAFGTSSDVEFNCTADNLAVEGAMCDLPAYKLMCFDIECKAGGEDELAFPVAERPEDLVIQISCLLYDLSTTALEHILLFSLGSCDLPESHLSDLASRGLPAPVVLEFDSEFEMLLAFMTFVKQYGPEFVTGYNIINFDWPFVLTKLTEIYKVPLDGYGRMNGRGVFRVWDIGQSHFQKRSKIKVNGMVNIDMYGIITDKVKLSSYKLNAVAEAVLKDKKKDLSYRDIPAYYASGPAQRGVIGEYCVQDSLLVGQLFFKFLPHLELSAVARLAGINITRTIYDGQQIRVFTCLLRLAGQKGFILPDTQGRFRGLDKEAPKRPAVPRGEGERPGDGNGDEDKDDDEDGDEDGDEREEVARETGGRHVGYQGARVLDPTSGFHVDPVVVFDFASLYPSIIQAHNLCFSTLSLRPEAVAHLEADRDYLEIEVGGRRLFFVKAHVRESLLSILLRDWLAMRKQIRSRIPQSTPEEAVLLDKQQAAIKVVCNSVYGFTGVQHGLLPCLHVAATVTTIGREMLLATRAYVHARWAEFDQLLADFPEAAGMRAPGPYSMRIIYGDTDSIFVLCRGLTAAGLVAMGDKMASHISRALFLPPIKLECEKTFTKLLLIAKKKYIGVICGGKMLIKGVDLVRKNNCAFINRTSRALVDLLFYDDTVSGAAAALAERPAEEWLARPLPEGLQAFGAVLVDAHRRITDPERDIQDFVLTAELSRHPRAYTNKRLAHLTVYYKLMARRAQVPSIKDRIPYVIVAQTREVEETVARLAALRELDAAAPGDEPAPPAALPSPAKRPRETPSHADPPGGASKPRKLLVSELAEDPGYAIARGVPLNTDYYFSHLLGAACVTFKALFGNNAKITESLLKRFIPETWHPPDDVAARLRAAGFGPAGAGATAEETRRMLHRAFDTLA SEQ ID NO: 31 =predicted sequence for helicase/primase complex encoded by UL5MAASGGEGSRDVRAPGPPPQQPGARPAVRFRDEAFLNFTSMHGVQPIIARIRELSQQQLDVTQVPRLQWFRDVAALEVPTGLPLREFPFAAYLITGNAGSGKSTCVQTLNEVLDCVVTGATRIAAQNMYVKLSGAFLSRPINTIFHEFGFRGNHVQAQLGQHPYTLASSPASLEDLQRRDLTYYWEVILDITKRALAAHGGEDARNEFHALTALEQTLGLGQGALTRLASVTHGALPAFTRSNIIVIDEAGLLGRHLLTTVVYCWWMINALYHTPQYAGRLRPVLVCVGSPTQTASLESTFEHQKLRCSVRQSENVLTYLICNRTLREYTRLSHSWAIFINNKRCVEHEFGNLMKVLEYGLPITEEHMQFVDRFVVPESYITNPANLPGWTRLFSSHKEVSAYMAKLHAYLKVTREGEFVVFTLPVLTFVSVKEFDEYRRLTQQPTLTMEKWITANASRITNYSQSQDQDAGHVRCEVHSKQQLVVARNDITYVLNSQVAVTARLRKMVFGFDGTFRTFEAVLRDDSFVKTQGETSVEFAYRFLSRLMFGGLIHFYNFLQRPGLDATQRTLAYGRLGELTAELLSLRRDAAGASATRAADTSDRSPGERAFNFKHLGPRDGGPDDFPDDDLDVIFAGLDEQQLDVFYCHYALEEPETTAAVHAQFGLLKRAFLGRYLILRELFGEVFESAPFS TYVDNVIFRGCELLTGSPRGGLMSVALQTDNYTLMGYTYTRVFAFAEELRRRHATAGVAEFLEESPLPYIVLRDQHGFMSVVNTNISEFVESIDSTELAMAINADYGISSKLAMTITRSQGLSLDKVAICFTPGNLRLNSAYVAMSRTTSSEFLHMNLNPLRERHERDDVISEHILSALRDPNVVIVY SEQ ID NO: 32 =predicted sequence for helicase/primase complex encoded by UL8MEAPGIVWVEESVSAITLYAVWLPPRTRDCLHALLYLVCRDAAGEARARFAEVSVGSSDLQDFYGSPDVSAPGAVAAARAATAPAASPLEPLGDPTLWRALYACVLAALERQTGRWALFVPLRLGWDPQTGLVVRVERASWGPPAAPRAALLDVEAKVDVDPLALSARVAEHPGARLAWARLAAIRDSPQCASSASLAVTITTRTARFAREYTTLAFPPTRKEGAFADLVEVCEVGLRPRGHPQRVTARVLLPRGYDYFVSAGDGFSAPALVALFRQWHTTVHAAPGALAPVFAFLGPGFEVRGGPVQYFAVLGFPGWPTFTVPAAAAAESARDLVRGAAATHAACLGAWPAVGARVVLPPRAWPAVASEAAGRLLPAFREAVARWHPTATTIQLLDPPAAVGPVWTARFCFSGLQAQLLAALAGLGEAGLPEARGRAGLERLDALVAAAPSEPWARAVLERLVPDACDACPALRQLLGGVMAAVCLQIEQTASSVKFAVCGGTGAAFWGLFNVDPGDADAAHGAIQDARRALEASVRAVLSANGIRPRLAPSLAPEGVYTHVVTWSQTGAWFWNSRDDTDFLQGFPLRGAAYAAAAEVMRDALRRILRRPAAGPPEEAVCAARGVMEDACDRFVLDAFGRRLDAEYWSVLTPPGEADDPLPQTAFRGGALLDAEQYWRRVVRVCPGGGESVGVPVDLYPRPLVLPPVDCAHHLREILREIQLVFTGVLEGVWGEGGSFVYPFDEKIRFLFPSEQ ID NO: 33 = predicted sequence for unknown protein encoded by UL15.5MDGAVRRTADLFLPDSFMQEIIGGQARETGDDRPVLTKSAGERFLLYRPSTTTNSGLMAPELYVYVDPAFTANTRAS GTGIAVVGRYRDDFIIFALEHFFLRALTGSAPADIARCVVHSLAQVLALHPGAFRSVRVAVEGNSSQDSAVAIATHVHTEMHRILASAGANGPGPELLFYHCEPPGGAVLYPFFLLNKQKTPAFEYFIKKFNSGGVMASQELVSVTVRLQTDPVEYLSEQLNNLIETVSPNTDVRMYSGKRNGAADDLMVAVIMAIYLAAPTGIPPAFFPITRTS SEQ ID NO: 34 =predicted sequence for cleavage and packaging protein encoded by UL32MATSAPGVPSSAAVREESPGSSWKEGAFERPYVAFDPDLLALNEALCAELLAACHVVGVPPASALDEDVESDVAPAPPRPRGAAREASGGRGPGSARGPPADPTAEGLLDTGPFAAASVDTFALDRPCLVCRTIELYKQAYRLSPQWVADYAFLCAKCLGAPHCAASIFVAAFEFVYVMDHHFLRTKKATLVGSFARFALTINDIHRHFFLHCCFRTDGGVPGRHAQKQPRPTPSPGAAKVQYSNYSFLAQSATRALIGTLASGGDDGAGAGAGGGSGTQPSLTTALMNWKDCARLLDCTEGKRGGGDSCCTRAAARNGEFEAAAGALAQGGEPETWAYADLILLLLAGTPAVWESGPRLRAAADARRAAVSESWEAHRGARMRDAAPRFAQFAEPQPQPDLDLGPLMATVLKHGRGRGRTGGECLLCNLLLVRAYWLAMRRLRASVVRYSENNTSLFDCIVPVVDQLEADPEAQPGDGGRFVSLLRAAGPEAIFKHMFCDPMCAITEMEVDPWVLFGHPRADHRDELQLHKAKLACGNEFEGRVCIALRALIYTFKTYQVFVPKPTALATFVREAGALLRRHSISLLSLEHTLCTYVSEQ ID NO: 35 =predicted sequence for ICP1/2 fragment enoded by UL36.4.2MEYDSFDALLAARLESGQTLGPAGGREASLPEAPHALYRPTGQHVAVLAAATHRTPAARVTAMDLVLAAVLLGAPVVVALRNTTAFSRESELELCLTLFDSRPGGPDAALRDVVSSDIETWAVGLLHTDLNPIENACLAAQLPRLSALIAERPLADGPPCLVLVDISMTPVAVLWEAPEPPGPPDVRFVGSEATEELPFVATAGDVLAASAADADPFFARAILGRPFDASLLTGELFPGHPVYQRPLADEAGPSAPTAARDPRDLAGGDGGSGPEDPAAPPARQADPGVLAPTLLTDATTGEPVPPRMWAWIHGLEELASDDAGGPT SEQ ID NO: 36 =predicted sequence for ICP27 encoded by UL54MATDIDMLIDLGLDLSDSELEEDALERDEEGRRDDPESDSSGECSSSDEDMEDPCGDGGAEAIDAAIPKGPPARPEDAGTPEASTPRPAARRGADDPPPATTGVWSRLGTRRSASPREPHGGKVARIQPPSTKAPHPRGGRRGRRRGRGRYGPGGADSTPKPRRRVSRNAHNQGGRHPASARTDGPGATHGEARRGGEQLDVSGGPRPRGTRQAPPPLMALSLTPPHADGRAPVPERKAPSADTIDPAVRAVLRSISERAAVERISESFGRSALVMQDPFGGMPFPAANSPWAPVLATQAGGFDAETRRVSWETLVAHGPSLYRTFAANPRAASTAKAMRDCVLRQENLIEALASADETLAWCKMCIHHNLPLRPQDPIIGTAAAVLENLATRLRPFLQCYLKARGLCGLDDLCSRRRLSDIKDIASFVLVILARLANRVERGVSEIDYTTVGVGAGETMHFYIPGACMAGLIEILDTHRQECSSRVCELTASHTIAPLYVHGKYFYCNSLF SEQ ID NO: 37 =virion protein encoded by UL49.5MTGKPARLGRWVVLLFVALVAGVPGEPPNAAGARGVIGDAQCRGDSAGVVSVPGVLVPFYLGMTSMGVCMIAHVYQICQRALAAGSA SEQ ID NO: 38 = gG2 encoded by US4NRWGSGVPGPINPPNSDVVFPGGSPVAQYCYAYPRLDDPGPLGSADAGRQDLPRRVVRHEPLGRSFLTGGLVLLAPPVRGFGAPNATYAARVTYYRLTRACRQPILLRQYGGCRGGEPPSPKTCGSYTYTYQGGGPPTRYALVNASLLVPIWDRAAETFEYQIELGGELHVGLLWVEVGGEGPGPTAPPQAARAEGGPCVPPVPAGRPWRSVPPVWYSAPNPGFRGLRFRERCLPPQTPAAPSDLPRVAFAPQSLLVGITGRTFIRMARPTEDVGVLPPHWAPGALDDGPYAPFPPRPRFRR SEQ ID NO: 39 =nucleotide sequence forRS1 (ICP4), full-lengthATGTCGTACTACCATCACCATCACCATCACAGTGCCGAACAGCGTAAAAAGAAAAAAACCACCACCACGACCCAAGGACGTGGAGCTGAAGTTGCTATGGCGGATGAGGATGGAGGCCGCTTGAGAGCTGCTGCTGAGACTACTGGAGGACCTGGATCACCGGACCCTGCCGATGGACCCCCCCCTACACCAAACCCCGATCGTAGACCGGCTGCTAGACCTGGATTCGGATGGCATGGAGGACCCGAGGAAAACGAGGACGAGGCGGACGACGCCGCTGCCGACGCCGACGCCGATGAGGCTGCCCCTGCTTCTGGAGAGGCGGTAGACGAACCTGCTGCCGATGGAGTTGTTAGCCCTAGGCAATTGGCTTTGTTGGCGAGCATGGTAGACGAGGCTGTGAGAACAATCCCTTCCCCTCCCCCTGAACGTGATGGAGCACAAGAGGAGGCGGCTAGGAGTCCCTCACCACCCCGTACACCTTCTATGAGAGCGGATTACGGCGAGGAAAACGACGACGACGACGATGATGATGACGACGATGATCGTGATGCCGGACGCTGGGTTAGGGGACCTGAAACCACTTCTGCTGTCCGTGGAGCATACCCCGATCCTATGGCGAGTTTGAGCCCTAGACCACCTGCCCCGAGGAGACACCACCACCACCACCATCATAGGCGTAGACGTGCTCCTAGACGTCGTTCTGCCGCTAGTGACTCTTCCAAATCTGGCTCTTCTTCATCTGCCTCTTCCGCTTCATCTTCGGCCTCATCGTCCTCTTCGGCATCCGCTTCGAGTAGTGATGATGATGATGACGACGACGCTGCTAGAGCCCCCGCTTCTGCTGCCGACCACGCTGCTGGCGGAACTTTGGGAGCCGACGACGAGGAGGCGGGAGTTCCTGCTCGTGCCCCGGGAGCTGCTCCGAGGCCTTCTCCACCCCGTGCTGAACCTGCTCCGGCTAGAACACCGGCCGCTACTGCTGGTAGACTGGAGCGTAGACGTGCCCGTGCTGCTGTGGCTGGTAGAGATGCTACTGGCCGCTTCACTGCTGGCCGTCCTAGACGTGTTGAACTGGACGCCGATGCTGCTTCTGGTGCTTTCTACGCCCGTTACCGTGATGGTTACGTGTCTGGTGAACCTTGGCCTGGCGCTGGTCCACCTCCGCCCGGACGTGTACTCTACGGTGGATTGGGCGATTCTCGCCCTGGTCTGTGGGGCGCTCCGGAGGCTGAGGAGGCTAGAGCCCGTTTCGAGGCTTCTGGTGCCCCTGCTCCTGTTTGGGCTCCTGAATTGGGCGACGCTGCTCAACAATACGCCCTCATCACACGCTTGCTGTACACTCCCGACGCCGAGGCTATGGGATGGCTCCAAAACCCTAGAGTTGCCCCTGGTGATGTTGCTCTGGATCAGGCTTGTTTCCGTATCTCCGGCGCTGCTCGTAACTCTTCTTCGTTCATCTCCGGTTCTGTGGCTAGAGCTGTGCCTCACTTGGGATACGCCATGGCCGCTGGACGTTTCGGCTGGGGACTGGCTCATGTTGCTGCCGCTGTAGCAATGTCTAGACGCTACGACCGTGCTCAAAAAGGATTCTTGCTCACGTCACTGAGGCGTGCTTACGCCCCTTTGTTGGCCCGTGAAAACGCTGCCCTCACTGGCGCCCGTACCCCCGATGACGGTGGCGACGCCAACCGCCACGATGGTGATGATGCTAGAGGCAAACCCGCTGCCGCTGCTGCTCCTTTGCCCTCTGCCGCCGCTTCCCCTGCCGATGAACGTGCTGTTCCTGCCGGTTACGGTGCCGCTGGTGTGTTGGCTGCTTTGGGACGCTTGAGTGCTGCCCCGGCTAGTGCCCCCGCTGGTGCCGATGACGATGACGATGACGATGGTGCTGGCGGAGGCGGTGGCGGTAGACGTGCTGAGGCTGGACGTGTTGCTGTTGAATGCCTGGCTGCCTGTAGAGGAATCTTGGAGGCTCTGGCCGAGGGATTCGACGGAGACTTGGCGGCTGTACCGGGACTGGCGGGAGCGAGGCCTGCCGCTCCACCTCGCCCCGGTCCTGCTGGTGCTGCCGCTCCTCCTCATGCCGACGCTCCTAGACTCCGTGCTTGGCTCCGTGAACTCCGTTTCGTTCGTGACGCTTTGGTTCTGATGAGACTGAGAGGCGACTTGAGAGTGGCTGGAGGATCCGAGGCTGCTGTTGCTGCTGTCCGTGCTGTTTCTTTGGTTGCTGGTGCTTTGGGCCCTGCTTTGCCGAGATCTCCCCGTTTGTTGTCGAGTGCCGCCGCTGCTGCCGCCGATTTGTTGTTCCAAAACCAATCCCTCCGCCCTCTGCTCGCCGACACTGTTGCCGCTGCCGATTCTCTGGCTGCTCCGGCTTCTGCCCCACGTGAAGCTCGTAAACGTAAATCACCCGCTCCGGCTCGTGCTCCCCCTGGTGGCGCCCCTAGACCCCCTAAAAAATCCCGTGCCGATGCCCCTAGACCTGCTGCTGCTCCCCCCGCTGGTGCTGCTCCCCCCGCTCCCCCTACTCCCCCCCCACGCCCACCTCGTCCCGCTGCCCTCACACGCCGTCCTGCTGAGGGACCCGATCCACAAGGCGGCTGGCGTAGACAACCTCCTGGCCCATCCCATACACCGGCACCATCTGCCGCTGCTTTGGAGGCTTACTGTGCTCCTCGTGCTGTGGCTGAACTCACCGATCATCCGCTGTTCCCTGCTCCCTGGCGTCCCGCCCTCATGTTCGATCCTAGAGCTTTGGCTTCCTTGGCCGCTCGTTGTGCTGCCCCTCCCCCTGGCGGTGCTCCGGCTGCTTTCGGTCCTCTCCGTGCCTCTGGTCCACTCCGCCGTGCCGCTGCCTGGATGAGACAAGTTCCCGACCCTGAGGATGTTAGAGTTGTGATCTTGTACTCGCCCTTGCCTGGCGAGGATTTGGCCGCTGGTAGAGCTGGCGGTGGCCCCCCTCCTGAATGGTCTGCTGAACGTGGTGGTTTGTCTTGCTTGTTGGCCGCCCTGGGAAACCGTCTGTGTGGTCCTGCTACTGCTGCTTGGGCTGGAAACTGGACTGGCGCTCCCGATGTTTCTGCTCTCGGTGCTCAAGGAGTTTTGCTGCTCTCTACTCGTGACTTGGCATTCGCTGGAGCTGTTGAATTCCTGGGACTCTTGGCTGGCGCTTGTGATAGGAGACTCATCGTCGTAAACGCTGTGAGAGCTGCCGATTGGCCTGCCGATGGTCCTGTTGTGTCTCGTCAACACGCTTACTTGGCTTGTGAAGTGTTGCCCGCTGTCCAATGTGCTGTTCGCTGGCCTGCTGCTCGTGATCTGAGGCGTACTGTTCTGGCTAGTGGTCGTGTTTTCGGACCTGGTGTTTTCGCTCGTGTCGAAGCTGCTCACGCTAGACTGTACCCCGATGCCCCACCCCTCCGTTTGTGTCGTGGAGCAAACGTTCGCTACCGTGTCCGTACTCGTTTCGGACCCGATACTCTGGTTCCAATGTCCCCTCGTGAATACCGTCGTGCTGTTCTGCCTGCCCTCGATGGACGTGCTGCCGCTTCTGGCGCTGGTGACGCTATGGCTCCTGGCGCTCCGGACTTCTGTGAGGATGAGGCTCACTCACATCGTGCCTGTGCCCGCTGGGGACTGGGCGCTCCATTGAGGCCTGTATACGTGGCACTGGGCCGTGATGCTGTTAGAGGCGGACCCGCTGAATTGAGAGGCCCTCGTCGTGAATTCTGTGCTAGGGCTCTGCTCGAACCCGATGGAGATGCTCCTCCTTTGGTACTCCGTGACGACGCCGATGCTGGTCCTCCCCCACAAATTCGCTGGGCTAGTGCTGCTGGACGTGCTGGTACTGTATTGGCTGCTGCTGGCGGTGGCGTTGAAGTTGTTGGTACTGCCGCTGGACTCGCTACACCTCCCCGCCGTGAACCTGTAGACATGGATGCTGAACTCGAGGATGATGACGACGGATTGTTCGGAGAGTAATAGSEQ ID NO: 40 = US6ΔTMRATGAAGTTCCTCGTGAACGTGGCCCTGGTGTTCATGGTGGTGTACATCAGCTACATCTACGCCAACCGTTGGAAGTACGCTCTGGCTGACCCATCCCTGAAGATGGCTGACCCCAACCGTTTCCGTGGCAAGAACCTGCCCGTGCTGGACCAGCTGACCGACCCCCCTGGCGTGAAGCGTGTGTACCACATCCAGCCATCCCTCGAAGACCCCTTCCAGCCCCCCTCCATCCCCATCACCGTGTACTACGCTGTGCTGGAACGCGCTTGCCGTTCCGTGCTGCTGCACGCTCCTTCCGAGGCTCCCCAGATCGTGCGTGGTGCTTCCGACGAGGCTCGCAAGCACACCTACAACCTGACTATCGCTTGGTACAGGATGGGTGACAACTGCGCTATCCCTATCACCGTCATGGAATACACCGAGTGCCCCTACAACAAGTCCCTGGGCGTGTGCCCTATCCGTACCCAGCCCCGTTGGTCCTACTACGACTCCTTCAGCGCTGTGTCCGAGGACAACCTGGGTTTCCTGATGCACGCTCCCGCTTTCGAGACTGCTGGCACCTACCTGCGTCTGGTCAAGATCAACGACTGGACCGAGATCACCCAGTTCATCCTGGAACACCGTGCTCGTGCTTCGTGCAAGTACGCCCTGCCCCTGCGTATCCCTCCTGCTGCTTGCCTGACCTCCAAGGCTTACCAGCAGGGCGTGACCGTGGACTCCATCGGCATGCTGCCCCGTTTCATCCCCGAGAACCAGCGTACCGTGGCTCTGTACTCTCTGAAGATCGCTGGCTGGCACGGTCCTAAGCCCCCCTACACCTCCACTCTGCTGCCCCCTGAGCTGTCCGACACCACCAACGCTACTCAGCCCGAGTTGGTGCCTGAGGACCCCGAGGACTCCGCTCTGTTGGAGGACCCCGCTGGAACCGTGTCCTCCCAGATCCCCCCCAACTGGCACATCCCTTCCATCCAGGACGTGGCCCCTCACCACGCTCCAGCTGCTCCCTCCAACCCCCGTCGTCGTGCTCAGATGGCTCCCAAGCGTCTGCGTCTGCCCCACATCCGTGACGACGACGCTCCTCCATCCCACCAGCCCCTGTTCTACCACCACCACCATCACCACTAATAA SEQ ID NO: 41 =nucleotide sequence for RL1 (ICP34.5)ATGTCTCGTCGTCGTGGTCCTCGTCGTCGTGGTCCTCGTCGTCGTCCGCGTCCGGGTGCGCCGGCGGTACCACGCCCGGGTGCGCCGGCAGTGCCGCGTCCAGGCGCACTGCCTACCGCGGACTCTCAAATGGTGCCGGCGTATGATTCTGGTACTGCCGTCGAATCTGCTCCGGCAGCGAGCTCCCTGCTGCGTCGTTGGCTGCTGGTCCCTCAGGCGGACGATTCCGATGACGCAGACTACGCGGGCAACGACGACGCGGAGTGGGCTAACAGCCCGCCAAGCGAGGGTGGTGGCAAAGCGCCGGAGGCTCCGCACGCAGCGCCTGCCGCAGCGTGCCCGCCTCCGCCTCCTCGTAAAGAACGTGGCCCTCAACGTCCTCTGCCGCCGCACCTGGCTCTGCGTCTGCGTACTACCACTGAGTACCTGGCGCGTCTGTCTCTGCGTCGTCGCCGTCCGCCGGCTAGCCCGCCGGCCGATGCACCGCGTGGCAAAGTGTGCTTCTCTCCACGTGTTCAAGTTCGTCACCTGGTGGCTTGGGAAACGGCTGCCCGTCTGGCTCGCCGTGGCAGCTGGGCACGTGAGCGCGCAGACCGTGACCGCTTCCGTCGCCGTGTGGCGGCTGCTGAAGCCGTTATCGGCCCGTGCCTGGAACCTGAGGCTCGCGCTCGCGCGCGTGCGCGCGCTCGTGCCCACGAAGATGGCGGTCCAGCAGAGGAAGAAGAGGCAGCTGCAGCAGCGCGCGGTAGCTCCGCGGCTGCGGGTCCAGGTCGTCGTGCCGTA SEQ ID NO: 42 = nucleotide seqeunce for RL2 (ICP0)ATGTCGTACTACCATCACCATCACCATCACATGGAGCCACGTCCTGGTACTTCTTCTCGCGCTGATCCTGGTCCTGAACGTCCGCCACGCCAGACTCCGGGCACCCAGCCGGCCGCCCCTCACGCTTGGGGCATGCTGAACGATATGCAGTGGCTGGCGTCCTCTGATTCCGAAGAGGAGACTGAGGTTGGTATCAGCGATGATGATCTGCACCGCGACTCTACCAGCGAAGCAGGTTCCACTGACACCGAAATGTTTGAAGCGGGCCTGATGGATGCCGCGACCCCGCCGGCTCGTCCGCCGGCTGAACGTCAGGGTAGCCCTACGCCTGCGGATGCGCAAGGCTCTTGTGGTGGTGGTCCAGTAGGCGAAGAGGAGGCTGAGGCCGGTGGCGGCGGTGATGTGTGTGCGGTTTGTACCGATGAAATCGCACCGCCGCTGCGTTGTCAGTCTTTCCCGTGCCTGCACCCGTTTTGCATTCCGTGCATGAAAACCTGGATCCCGCTGCGCAACACTTGCCCGCTGTGCAACACTCCGGTTGCTTATCTGATCGTTGGTGTAACCGCATCTGGTTCCTTTTCTACCATCCCGATTGTCAACGACCCACGTACGCGTGTTGAGGCGGAGGCGGCTGTACGTGCGGGCACCGCGGTGGACTTTATCTGGACCGGTAACCCGCGCACCGCGCCACGCTCCCTGTCTCTGGGTGGCCATACCGTTCGTGCTCTGAGCCCGACCCCACCTTGGCCAGGCACCGATGACGAAGACGACGATCTGGCTGACGTTGACTATGTTCCGCCGGCACCGCGTCGCGCACCACGCCGTGGTGGCGGTGGCGCCGGTGCGACGCGCGGTACCTCCCAGCCGGCAGCAACTCGCCCAGCACCGCCGGGTGCCCCGCGTTCTAGCAGCTCCGGTGGCGCACCGCTGCGTGCTGGCGTGGGTTCTGGTTCCGGTGGTGGTCCGGCCGTGGCGGCTGTCGTCCCGCGTGTGGCTTCTCTGCCACCGGCAGCTGGTGGCGGTCGTGCTCAAGCTCGTCGTGTCGGCGAGGACGCAGCGGCTGCTGAGGGCCGTACTCCACCGGCCCGTCAACCGCGCGCAGCACAGGAACCGCCGATCGTGATCTCCGATTCCCCGCCACCGAGCCCGCGTCGCCCGGCGGGTCCGGGTCCGCTGTCTTTTGTATCCTCCAGCTCTGCTCAGGTAAGCAGCGGTCCTGGCGGTGGCGGCCTGCCACAGTCCTCTGGTCGTGCTGCTCGTCCTCGTGCGGCGGTTGCTCCTCGTGTACGTTCTCCGCCACGCGCTGCTGCCGCGCCGGTCGTTTCTGCCTCTGCTGACGCGGCAGGTCCGGCTCCGCCTGCAGTTCCGGTTGATGCACACCGTGCACCGCGCTCTCGTATGACCCAGGCGCAGACTGATACCCAGGCACAATCCCTGGGTCGCGCGGGTGCGACTGACGCTCGTGGTAGCGGTGGTCCGGGCGCTGAAGGTGGCCCGGGTGTTCCACGCGGTACTAACACTCCGGGCGCTGCGCCACACGCGGCTGAAGGTGCGGCTGCACGTCCGCGTAAACGTCGTGGTTCCGACAGCGGTCCGGCTGCAAGCAGCAGCGCGAGCTCTTCCGCTGCGCCTCGCAGCCCGCTGGCGCCGCAGGGTGTTGGCGCCAAGCGTGCTGCTCCGCGTCGTGCACCGGACTCCGATTCTGGCGACCGCGGTCACGGCCCGCTGGCCCCTGCTAGCGCAGGCGCTGCGCCGCCATCCGCCAGCCCGTCTTCTCAGGCAGCTGTGGCTGCGGCGTCCTCTTCTTCCGCTAGCAGCTCTTCCGCCTCTTCTAGCAGCGCGTCCTCTAGCAGCGCATCTTCCTCTTCTGCTTCTTCTTCTAGCGCTTCTAGCTCTTCCGCGTCCTCTTCCGCTGGCGGTGCAGGCGGCTCTGTTGCTTCCGCCAGCGGCGCAGGTGAGCGTCGTGAAACGAGCCTGGGCCCACGTGCTGCTGCACCGCGTGGCCCGCGTAAGTGTGCGCGCAAGACCCGCCACGCTGAAGGCGGTCCGGAGCCGGGTGCGCGTGATCCGGCTCCGGGTCTGACCCGTTACCTGCCGATTGCGGGTGTGTCCTCCGTTGTGGCACTGGCGCCGTATGTGAACAAAACTGTCACGGGCGATTGCCTGCCTGTTCTGGACATGGAAACCGGTCATATCGGCGCTTACGTCGTTCTGGTTGACCAAACCGGCAACGTGGCGGATCTGCTGCGTGCGGCCGCTCCGGCTTGGTCCCGTCGTACCCTGCTGCCGGAACATGCTCGCAACTGTGTACGCCCACCGGATTACCCAACCCCGCCGGCCTCCGAGTGGAACTCCCTGTGGATGACCCCGGTTGGTAACATGCTGTTCGACCAGGGCACGCTGGTTGGTGCTCTGGACTTTCACGGCCTGCGCTCCCGTCACCCGTGGTCCCGTGAGCAAGGCGCTCCGGCCCCTGCGGGCGATGCCCCGGCTGGCCACGGCGAGAGTACTAGAGGATCATAASEQ ID NO: 43 = nucleotide sequence for UL36.3.4.1ATGTCGTACTACCATCACCATCACCATCACGCCGCTCAACGTGCTAGGGGATCCTCTGAACGCTGGGCTGCTGGTGTCGAGGCTGCTTTGGATAGAGTGGAGAACCGTGCCGAATTCGATGTTGTCGAGCTGAGGAGACTCCAAGCTTTGGCTGGTACTCACGGCTACAACCCTCGTGATTTCCGTAAACGTGCCGAACAGGCTTTGGCGGCAAACGCTGAGGCCGTAACATTGGCTCTGGACACTGCCTTCGCTTTCAACCCATACACGCCCGAAAACCAACGTCATCCTATGCTCCCACCTCTCGCTGCTATTCACCGCCTGGGATGGAGCGCTGCTTTCCATGCTGCTGCTGAAACTTACGCCGACATGTTCCGTGTCGATGCCGAACCACTGGCTAGACTGCTCCGTATCGCTGAGGGACTGCTGGAGATGGCTCAAGCTGGCGACGGATTCATCGATTACCATGAGGCTGTCGGTAGACTGGCCGATGATATGACTTCTGTGCCCGGATTGAGGCGCTACGTTCCTTTCTTCCAACATGGCTACGCCGATTACGTGGAACTGAGAGATCGCCTGGATGCTATTAGGGCCGACGTCCATAGAGCACTCGGTGGTGTTCCGCTGGATTTGGCGGCTGCTGCCGAACAAATTTCCGCTGCTCGTAACGATCCTGAGGCTACTGCTGAATTGGTCCGTACTGGTGTAACATTGCCTTGCCCTAGTGAGGACGCTCTCGTGGCTTGTGCTGCTGCCCTGGAGAGAGTCGATCAATCTCCCGTGAAAAACACGGCTTACGCCGAATACGTTGCCTTCGTGACCCGTCAAGACACTGCTGAGACTAAAGACGCTGTGGTCCGTGCTAAACAACAACGTGCTGAGGCCACTGAACGTGTTATGGCTGGCCTGAGAGAGGCTCTGGCTGCTAGAGAACGTCGTGCTCAAATTGAGGCTGAGGGATTGGCAAACCTGAAAACCATGCTCAAAGTCGTGGCTGTACCCGCTACTGTTGCTAAAACTCTCGACCAGGCTCGTAGTGTTGCCGAAATTGCCGATCAAGTCGAAGTGTTGCTGGATCAAACCGAAAAAACTCGTGAACTGGATGTGCCTGCTGTGATCTGGCTCGAACACGCCCAAAGAACATTCGAGACACACCCTTTGTCTGCCGCTCGTGGTGATGGTCCTGGACCCTTGGCTCGTCATGCTGGCCGCCTCGGTGCCCTCTTCGATACTCGTCGTAGAGTAGACGCCTTGAGGAGATCCCTGGAGGAGGCTGAGGCTGAATGGGACGAAGTTTGGGGACGCTTCGGTAGAGTGAGGGGCGGAGCGTGGAAATCTCCGGAGGGATTCCGTGCAATGCATGAGCAACTGAGGGCCCTCCAAGACACAACAAACACCGTGTCTGGCCTGAGGGCTCAACCTGCTTACGAACGCTTGTCTGCTCGCTACCAAGGAGTACTCGGAGCGAAAGGCGCTGAGAGAGCTGAGGCTGTTGAGGAACTCGGTGCTCGTGTCACTAAACACACCGCTCTGTGTGCTAGGCTGAGAGATGAGGTCGTCCGTAGAGTGCCTTGGGAAATGAACTTCGATGCTCTGGGAGGATTGTTGGCTGAGTTCGATGCCGCTGCTGCCGATTTGGCACCTTGGGCTGTAGAGGAATTCCGTGGTGCTAGAGAACTCATTCAATACCGTATGGGCCTGTACTCTGCCTACGCTAGAGCTGGAGGACAAACTGGTGCTGGAGCTGAATCTGCTCCTGCTCCTTTGCTCGTGGATCTGAGGGCTTTGGATGCTCGTGCTCGTGCTTCTTCTTCCCCTGAGGGACATGAAGTGGACCCACAACTGCTGAGGAGGCGTGGAGAGGCTTACTTGAGAGCTGGCGGCGACCCTGGACCTCTCGTGCTCCGTGAAGCTGTTTCTGCTTTGGACCTGCCATTCGCCACATCTTTCTTGGCCCCCGATGGAACTCCCCTCCAATACGCTTTGTGCTTCCCTGCCGTAACGGACAAACTCGGAGCTTTGCTCATGAGGCCCGAGGCCGCTTGTGTTAGACCTCCTTTGCCTACCGATGTGCTGGAATCTGCCCCAACTGTGACTGCCATGTACGTACTCACTGTGGTCAACCGCCTCCAACTGGCATTGAGTGATGCTCAAGCGGCAAACTTCCAACTGTTCGGTCGTTTCGTTCGTCATAGGCAGGCAACCTGGGGAGCGTCAATGGATGCCGCCGCTGAATTGTACGTTGCCCTGGTGGCTACAACTCTCACACGTGAATTCGGTTGTCGCTGGGCACAATTGGGATGGGCTAGTGGAGCTGCTGCTCCTAGACCCCCACCTGGACCCCGTGGCTCACAACGTCACTGTGTGGCATTCAACGAGAACGATGTCCTCGTCGCTTTGGTTGCCGGTGTTCCCGAACACATCTACAACTTCTGGCGCCTGGACTTGGTCCGTCAACACGAGTACATGCACCTCACACTGGAGCGTGCCTTCGAGGATGCTGCCGAGTCTATGCTCTTCGTTCAACGCCTCACTCCACATCCCGACGCTCGTATTAGAGTTCTGCCGACCTTCTTGGATGGTGGTCCTCCTACACGTGGTCTGTTGTTCGGAACCCGCTTGGCGGACTGGCGTCGTGGTAAACTGTCTGAAACCGACCCATTGGCCCCATGGAGATCTGCTTTGGAACTCGGAACCCAACGTCGTGACGTGCCTGCTTTGGGAAAACTGTCCCCTGCTCAAGCTTTGGCCGCTGTGTCGGTACTGGGCCGTATGTGCTTGCCCTCGGCTGCCTTGGCTGCTTTGTGGACCTGTATGTTCCCCGACGACTACACTGAATACGACTCATTCGACGCCCTCTTGGCGGCTCGCCTGGAATCGGGACAAACATTGGGACCTGCTGGCGGTAGAGAGGCTTCATTGTAATAG SEQ ID NO: 44 = nucleotide sequence for UL36.4.2.5ATGTCGTACTACCATCACCATCACCATCACGAATACGACTCCTTCGACGCTTTGTTGGCTGCTAGACTGGAATCTGGTCAAACCTTGGGACCCGCTGGCGGTAGAGAGGCTTCTTTGCCCGAGGCTCCTCATGCTTTGTACCGTCCAACCGGACAACATGTTGCTGTGTTGGCGGCTGCTACTCATAGAACCCCTGCTGCTCGTGTTACTGCTATGGACCTGGTCTTGGCGGCCGTTTTGCTGGGCGCTCCTGTGGTGGTCGCTCTGAGAAACACTACTGCCTTCTCCCGTGAATCCGAATTGGAACTGTGCCTCACCCTGTTCGATTCTCGTCCCGGCGGACCGGATGCTGCCCTGAGAGATGTGGTATCCTCCGACATTGAAACCTGGGCTGTGGGCTTGCTCCACACCGATTTGAACCCTATTGAGAACGCTTGCTTGGCGGCTCAACTGCCACGCTTGTCTGCCCTCATTGCTGAACGTCCTTTGGCCGATGGACCCCCTTGTTTGGTGTTGGTGGACATTTCGATGACACCTGTCGCTGTTTTGTGGGAGGCCCCTGAACCACCTGGCCCTCCCGATGTTCGTTTCGTCGGTAGCGAGGCCACTGAGGAATTGCCTTTCGTGGCTACTGCTGGTGATGTTTTGGCGGCGAGTGCTGCCGATGCCGATCCTTTCTTCGCTCGTGCTATCCTGGGCCGTCCTTTCGATGCTTCTCTGCTCACTGGTGAACTGTTCCCTGGTCACCCCGTTTACCAACGTCCCCTGGCGGATGAGGCTGGTCCTTCTGCTCCTACTGCCGCTCGTGATCCTAGAGATCTGGCTGGAGGCGACGGTGGATCCGGACCTGAGGATCCCGCTGCTCCACCTGCTAGACAGGCCGATCCTGGTGTTTTGGCTCCTACTCTGCTCACCGATGCTACTACTGGCGAACCTGTGCCACCCCGTATGTGGGCTTGGATTCATGGACTGGAGGAACTGGCTTCCGATGATGCCGGCGGTCCTACCCCAAACCCTGCCCCGGCTTTGCTGCCCCCTCCTGCTACGGATCAATCTGTCCCCACTTCCCAATACGCCCCTAGACCAATTGGCCCGGCTGCCACTGCTAGAGAAACTCGTCCTTCCGTTCCCCCTCAACAAAACACTGGTCGTGTCCCTGTGGCTCCACGTGATGACCCTAGACCTTCCCCCCCTACTCCTTCCCCCCCTGCCGATGCTGCTTTGCCACCTCCTGCCTTCTCTGGTTCTGCTGCTGCTTTCTCCGCTGCTGTTCCACGTGTTCGTCGTTCTAGGCGTACTCGTGCCAAATCCCGTGCCCCTCGTGCTTCTGCCCCACCCGAGGGATGGCGTCCCCCCGCTTTGCCTGCCCCTGTTGCTCCTGTGGCGGCTTCTGCTCGTCCCCCCGATCAACCTCCTACTCCCGAATCTGCTCCCCCGGCTTGGGTTTCCGCTCTGCCATTGCCACCCGGACCTGCTAGTGCTCGTGGTGCTTTCCCTGCTCCAACCTTGGCCCCTATTCCCCCACCCCCCGCTGAGGGAGCTGTTGTTCCCGGTGGTGATCGTAGACGTGGTCGCCGTCAAACAACTGCTGGACCATCCCCTACACCGCCACGTGGCCCGGCTGCTGGTCCTCCTCGTCGCCTCACTAGGCCTGCTGTTGCTAGTCTGTCCGCTTCTTTGAACTCTCTGCCTTCCCCCCGTGATCCTGCCGATCATGCTGCTGCCGTTTCTGCTGCCGCCGCTGCCGTACCACCTTCACCTGGACTGGCTCCCCCAACTTCTGCTGTCCAAACCTCTCCTCCTCCCTTGGCGCCTGGTCCTGTTGCCCCATCTGAACCTTTGTGTGGCTGGGTTGTGCCTGGAGGCCCTGTTGCTAGACGTCCCCCACCCCAATCTCCGGCTACTAAACCGGCTGCTCGTACCCGTATTAGGGCTCGTTCTGTGCCCCAACCACCCTTGCCCCAACCTCCACTGCCTCAACCCCCCTTGCCTCAACCCCCTCTCCCCCAACCACCTCTGCCTCAACCTCCGCTGCCCCAACCTCCTTTGCCCCAACCTCCTTTGCCCCAACCTCCTTTGCCCCAACCTCCGCTGCCCCAACCTCCGCTGCCACCTGTTACTCGTACACTCACTCCCCAATCTCGTGACTCTGTGCCTACACCTGAGTCTCCAACTCACACAAACACCCACTTGCCCGTTAGTGCTGTGACTTCTTGGGCTTCGTCCCTGGCTCTCCATGTGGATTCTGCCCCTCCCCCTGCTTCATTGCTCCAAACTCTCCACATTTCCTCCGATGATGAACACTCCGACGCCGACTCACTCCGCTTCTCCGATTCCGATGACACTGAGGCTCTCGATCCTTTGCCTCCTGAACCTCACTTGCCACCTGCCGATGAACCCCCCGGACCTCTGGCTGCCGACCATCTCCAATCACCTCACTCACAATTCGGTCCTTTGCCCGTTCAAGCGAACGCTGTTCTGTCTCGTCGTTACGTGAGATCAACTGGCCGTTCTGCCTTGGCTGTGCTCATTAGAGCTTGTCGCCGTATCCAACAACAACTCCAGCGTACTAGGAGAGCACTCTTCCAACGCTCAAACGCCGTGCTCACATCACTCCACCATGTCCGTATGCTCTTGGGATAATAG SEQ ID NO: 45 =nucleotide sequence for US12 (ICP47)ATGTCTTGGGCTCTGAAAACCACCGACATGTTCCTGGACTCTTCTCGTTGCACCCACCGTACCTACGGTGACGTTTGCGCTGAAATCCACAAACGTGAACGTGAAGACCGTGAAGCTGCTCGTACCGCTGTTACCGACCCGGAACTGCCGCTGCTGTGCCCGCCGGACGTTCGTTCTGACCCGGCTTCTCGTAACCCGACCCAGCAGACCCGTGGTTGCGCTCGTTCTAACGAACGTCAGGACCGTGTTCTGGCTCCGTGA SEQ ID NO: 46 =nucleotide sequence for US4ATGAAGTTCCTCGTGAACGTGGCCCTGGTGTTCATGGTGGTGTACATCAGCTACATCTACGCTAACCGTTGGGGTTCCGGCGTGCCCGGTCCCATCAACCCCCCCAACTCCGACGTGGTGTTCCCCGGTGGTTCCCCCGTGGCTCAGTACTGCTACGCTTACCCCCGTCTGGACGACCCTGGTCCCCTGGGTTCTGCTGACGCTGGTCGTCAGGACCTGCCCCGTCGTGTCGTGCGTCACGAGCCCCTGGGTCGTAGCTTCCTGACCGGTGGCCTGGTGCTGTTGGCTCCCCCTGTGCGCGGTTTCGGTGCTCCCAACGCTACCTACGCTGCTCGTGTGACCTACTACCGTCTGACCCGTGCTTGCCGTCAGCCCATCCTGCTGCGTCAGTACGGTGGTTGCCGTGGTGGAGAGCCCCCATCCCCCAAGACCTGCGGTTCTTACACCTACACCTACCAGGGTGGTGGTCCCCCTACCCGTTACGCTCTGGTCAACGCTTCCCTGCTGGTGCCCATCTGGGACCGTGCTGCTGAGACTTTCGAGTACCAGATCGAGCTGGGTGGCGAGCTGCACGTGGGTCTGCTGTGGGTGGAAGTGGGTGGAGAGGGTCCCGGTCCTACCGCTCCTCCTCAGGCTGCTCGTGCTGAGGGTGGTCCTTGCGTGCCACCCGTGCCTGCTGGTCGTCCTTGGCGTTCCGTGCCCCCCGTGTGGTACTCCGCTCCCAACCCCGGTTTCCGCGGTCTGCGTTTCCGTGAGCGTTGCCTGCCTCCCCAGACCCCTGCTGCTCCTTCCGACCTGCCTCGTGTGGCTTTCGCTCCCCAGTCCCTGCTCGTGGGTATCACCGGTCGTACCTTCATCCGTATGGCTCGTCCCACCGAGGACGTGGGTGTCCTGCCTCCTCACTGGGCTCCAGGTGCTCTGGACGACGGTCCCTACGCTCCCTTCCCCCCTCGTCCCCGTTTCCGTCGTCACCACCACCATCACCACTAATAASEQ ID NO: 117 = RS1.2ATGTCGTACTACCATCACCATCACCATCACATGGTGCTGTACGGCGGGCTGGGCGACAGCCGCCCCGGCCTCTGGGGGGCGCCCGAGGCGGAGGAGGCGCGGGCCCGGTTCGAGGCCTCGGGCGCCCCGGCGCCCGTGTGGGCGCCCGAGCTGGGCGACGCGGCGCAGCAGTACGCCCTGATCACGCGGCTGCTGTACACGCCGGACGCGGAGGCGATGGGGTGGCTCCAGAACCCGCGCGTGGCGCCCGGGGACGTGGCGCTGGACCAGGCCTGCTTCCGGATCTCGGGCGCGGCGCGCAACAGCAGCTCCTTCATCTCCGGCAGCGTGGCGCGGGCCGTGCCCCACCTGGGGTACGCCATGGCGGCGGGCCGCTTCGGCTGGGGCCTGGCGCACGTGGCGGCCGCCGTGGCCATGAGCCGCCGCTACGACCGCGCGCAGAAGGGCTTCCTGCTGACCAGCCTGCGCCGCGCCTACGCGCCCCTGCTGGCGCGCGAGAACGCGGCGCTGACCGGGGCGCGGACCCCCGACGACGGCGGCGACGCCAACCGCCGCGACGGCGACGACGCCCGCGGGAAGCCCGCCGCCGCCGCCGCCCCGTTGCCGTCGGCGGCGGCGTCGCCGGCCGACGAGCGCGCGGTGCCCGCCGGCTACGGCGCCGCGGGGGTGCTCGCCGCCCTGGGGCGCCTGAGCGCCGCGCCCGCCTCCGCGCCGGCCGGGGCCGACGACGACGACGACGACGACGACGGCGCCGGCGGTGGTGGCGGTGGTGGCGGTGGTGGCGGCGGCCGGCGCGCGGAGGCGGGCCGCGTGGCCGTGGAGTGCCTGGCCGCCTGCCGCGGGATCCTGGAGGCGCTGGCGGAGGGCTTCGACGGCGACCTGGCGGCCGTGCCGGGGCTGGCCGGAGCCCGGCCCGCCGCGCCCCCGCGCCCGGGGCCCGCGGGCGCGGCCGCCCCGCCGCACGCCGACGCGCCCCGCCTGCGCGCCTGGCTGCGCGAGCTGCGGTTCGTGCGCGACGCGCTGGTGCTGATGCGCCTGCGCGGGGACCTGCGCGTGGCCGGCGGCAGCGAGGCCGCCGTGGCCGCCGTGCGCGCCGTGAGCCTGGTCGCCGGGGCCCTGGGCCCGGCGCTGCCGCGGAGCCCGCGCCTGCTGAGCTCCGCCGCCGCCGCCGCCGCGGACCTGCTCTTCCAGAACCAGAGCCTGAGTACTAGAGGATCATAASEQ ID NO: 118 = UL1 (cytoplasmic), gLfull lengthATGTCGTACTACCATCACCATCACCATCACATGGGGTTCGTCTGTCTGTTTGGGCTTGTCGTTATGGGAGCCTGGGGGGCGTGGGGTGGGTCACAGGCAACCGAATATGTTCTTCGTAGTGTTATTGCCAAAGAGGTGGGGGACATACTAAGAGTGCCTTGCATGCGGACCCCCGCGGACGATGTTTCTTGGCGCTACGAGGCCCCGTCCGTTATTGACTATGCCCGCATAGACGGAATATTTCTTCGCTATCACTGCCCGGGGTTGGACACGTTTTTGTGGGATAGGCACGCCCAGAGGGCGTATCTTGTTAACCCCTTTCTCTTTGCGGCGGGATTTTTGGAGGACTTGAGTCACTCTGTGTTTCCGGCCGACACCCAGGAAACAACGACGCGCCGGGCCCTTTATAAAGAGATACGCGATGCGTTGGGCAGTCGAAAACAGGCCGTCAGCCACGCACCCGTCAGGGCCGGGTGTGTAAACTTTGACTACTCACGCACTCGCCGCTGCGTCGGGCGACGCGATTTACGGCCTGCCAACACCACGTCAACGTGGGAACCGCCTGTGTCGTCGGACGATGAAGCGAGCTCGCAGTCGAAGCCCCTCGCCACCCAGCCGCCCGTCCTCGCCCTTTCGAACGCCCCCCCACGGCGGGTCTCCCCGACGCGAGGTCGGCGCCGGCATACTCGCCTCCGACGCAACTGA SEQ ID NO: 119 =UL1 (Secreted), gLfull length (preferred Ag)ATGAAGTTCCTCGTGAACGTGGCCCTGGTGTTCATGGTGGTGTACATCAGCTACATCTACGCCAACCGTTGGGGGTTCGTCTGTCTGTTTGGGCTTGTCGTTATGGGAGCCTGGGGGGCGTGGGGTGGGTCACAGGCAACCGAATATGTTCTTCGTAGTGTTATTGCCAAAGAGGTGGGGGACATACTAAGAGTGCCTTGCATGCGGACCCCCGCGGACGATGTTTCTTGGCGCTACGAGGCCCCGTCCGTTATTGACTATGCCCGCATAGACGGAATATTTCTTCGCTATCACTGCCCGGGGTTGGACACGTTTTTGTGGGATAGGCACGCCCAGAGGGCGTATCTTGTTAACCCCTTTCTCTTTGCGGCGGGATTTTTGGAGGACTTGAGTCACTCTGTGTTTCCGGCCGACACCCAGGAAACAACGACGCGCCGGGCCCTTTATAAAGAGATACGCGATGCGTTGGGCAGTCGAAAACAGGCCGTCAGCCACGCACCCGTCAGGGCCGGGTGTGTAAACTTTGACTACTCACGCACTCGCCGCTGCGTCGGGCGACGCGATTTACGGCCTGCCAACACCACGTCAACGTGGGAACCGCCTGTGTCGTCGGACGATGAAGCGAGCTCGCAGTCGAAGCCCCTCGCCACCCAGCCGCCCGTCCTCGCCCTTTCGAACGCCCCCCCACGGCGGGTCTCCCCGACGCGAGGTCGGCGCCGGCATACTCGCCTCCGACGCAACCATCACCATCACCATCACTGASEQ ID NO: 120 UL19 delta TEV VP5 full lengthATGTCGTACTACCATCACCATCACCATCACATGGCCGCTCCTGCCCGCGACCCCCCGGGTTACCGGTACGCCGCGGCCATGGTGCCCACCGGCTCCATCCTGAGTACGATCGAGGTGGCGTCCCACCGCAGACTCTTTGATTTTTTCGCCCGCGTGCGCTCCGACGAAAACAGCCTGTATGACGTAGAGTTTGACGCCCTGCTGGGGTCCTACTGCAACACCCTGTCGCTCGTGCGCTTTCTGGAGCTCGGCCTGTCCGTGGCGTGCGTGTGCACCAAGTTCCCGGAGCTGGCTTACATGAACGAAGGGCGTGTGCAGTTCGAGGTCCACCAGCCCCTCATCGCCCGCGACGGCCCGCACCCCGTCGAGCAGCCCGTGCATAATTACATGACGAAGGTCATCGACCGCCGGGCCCTGAACGCCGCCTTCAGCCTGGCCACCGAGGCCATTGCCCTGCTCACGGGGGAGGCCCTGGACGGGACGGGCATTAGCCTGCATCGCCAGCTGCGCGCCATCCAGCAGCTCGCGCGCAACGTCCAGGCCGTCCTGGGGGCGTTTGAGCGCGGCACGGCCGACCAGATGCTGCACGTGCTGTTGGAGAAGGCGCCTCCCCTGGCCCTGCTGTTGCCCATGCAACGATATCTCGACAACGGGCGCCTGGCGACCAGGGTTGCCCGGGCGACCCTGGTCGCCGAGCTGAAGCGGAGCTTTTGCGACACGAGCTTCTTCCTGGGCAAGGCGGGCCATCGCCGCGAGGCCATCGAGGCCTGGCTCGTGGACCTGACCACGGCGACGCAGCCGTCCGTGGCCGTGCCCCGCCTGACGCACGCCGACACGCGCGGGCGGCCGGTCGACGGGGTGCTGGTCACCACCGCCGCCATCAAACAGCGCCTCCTGCAGTCCTTCCTGAAGGTGGAGGACACCGAGGCCGACGTGCCGGTGACCTACGGCGAGATGGTCTTGAACGGGGCCAACCTCGTCACGGCGCTGGTGATGGGCAAGGCCGTGCGGAGCCTGGACGACGTGGGCCGCCACCTGCTGGAGATGCAGGAGGAGCAACTCGAGGCGAACCGGGAGACGCTGGATGAACTCGAAAGCGCCCCCCAGACAACGCGCGTGCGCGCGGATCTGGTGGCCATAGGCGACAGGCTGGTCTTCCTGGAGGCCCTGGAGAAGCGCATCTACGCCGCCACCAACGTGCCCTACCCCCTGGTGGGCGCCATGGACCTGACGTTCGTCCTGCCCCTGGGGCTGTTCAACCCGGCCATGGAGCGCTTCGCCGCGCACGCCGGGGACCTGGTGCCCGCCCCCGGCCACCCGGAGCCCCGCGCGTTCCCTCCCCGGCAGCTGTTTTTTTGGGGAAAGGACCACCAGGTTCTGCGGCTGTCCATGGAGAACGCGGTCGGGACCGTGTGTCATCCTTCGCTCATGAACATCGACGCGGCCGTCGGGGGCGTGAACCACGACCCCGTCGAGGCCGCGAATCCGTACGGGGCGTACGTCGCGGCCCCGGCCGGCCCCGGCGCGGACATGCAGCAGCGTTTTCTGAACGCCTGGCGGCAGCGCCTCGCCCACGGCCGGGTCCGGTGGGTCGCCGAGTGCCAGATGACCGCGGAGCAGTTCATGCAGCCCGACAACGCCAACCTGGCTCTGGAGCTGCACCCCGCGTTCGACTTCTTCGCGGGCGTGGCCGACGTCGAGCTTCCCGGCGGCGAAGTCCCCCCGGCCGGTCCGGGGGCGATCCAGGCCACCTGGCGCGTGGTCAACGGCAACCTGCCCCTGGCGCTGTGTCCGGTGGCGTTTCGTGACGCCCGGGGCCTGGAGCTCGGCGTTGGCCGCCACGCCATGGCGCCGGCTACCATAGCCGCCGTCCGCGGGGCGTTCGAGGACCGCAGCTACCCGGCGGTGTTCTACCTGCTGCAAGCCGCGATTCACGGCAGCGAGCACGTGTTCTGCGCCCTGGCGCGGCTCGTGACTCAGTGCATCACCAGCTACTGGAACAACACGCGATGCGCGGCGTTCGTGAACGACTACTCGCTGGTCTCGTACATCGTGACCTACCTCGGGGGCGACCTCCCCGAGGAGTGCATGGCCGTGTATCGGGACCTGGTGGCCCACGTCGAGGCCCTGGCCCAGCTGGTGGACGACTTTACCCTGCCGGGCCCGGAGCTGGGCGGGCAGGCTCAGGCCGAGCTGAATCACCTGATGCGCGACCCGGCGCTGCTGCCGCCCCTCGTGTGGGACTGCGACGGCCTTATGCGACACGCGGCCCTGGACCGCCACCGAGACTGCCGGATTGACGCGGGGGAGCACGAGCCCGTCTACGCGGCGGCGTGCAACGTGGCGACGGCCGACTTTAACCGCAACGACGGCCGGCTGCTGCACAACACCCAGGCCCGCGCGGCCGACGCCGCCGACGACCGGCCGCACCGGCCGGCCGACTGGACCGTCCACCACAAAATCTACTATTACGTGCTGGTGCCGGCCTTCTCGCGGGGGCGCTGCTGCACCGCGGGGGTCCGCTTCGACCGCGTGTACGCCACGCTGCAGAACATGGTGGTCCCGGAGATCGCCCCCGGCGAGGAGTGCCCGAGCGATCCCGTGACCGACCCCGCCCACCCGCTGCATCCCGCCAATCTGGTGGCCAACACGGTCAACGCCATGTTCCACAACGGGCGCGTCGTCGTCGACGGGCCCGCCATGCTCACGCTGCAGGTGCTGGCGCACAACATGGCCGAGCGCACGACGGCGCTGCTGTGCTCCGCGGCGCCCGACGCGGGCGCCAACACCGCGTCGACGGCCAACATGCGCATCTTCGACGGGGCGCTGCACGCCGGCGTGCTGCTCATGGCCCCCCAGCACCTGGACCACACCATCCAAAATGGCGAATACTTCTACGTCCTGCCCGTCCACGCGCTGTTTGCGGGCGCCGACCACGTGGCCAACGCGCCCAACTTCCCCCCGGCCCTGCGCGACCTGGCGCGCCACGTCCCCCTGGTCCCCCCGGCCCTGGGGGCCAACTACTTCTCCTCCATCCGCCAGCCCGTGGTGCAGCACGCCCGCGAGAGCGCGGCGGGGGAGAACGCGCTGACCTACGCGCTCATGGCGGGGTACTTCAAGATGAGCCCCGTGGCCCTGTATCACCAGCTCAAGACGGGCCTCCACCCCGGGTTCGGGTTCACCGTCGTGCGGCAGGACCGCTTCGTGACCGAGAACGTGCTGTTTTCCGAGCGCGCGTCGGAGGCGTACTTTCTGGGCCAGCTCCAGGTGGCCCGCCACGAAACGGGCGGGGGGGTCAGCTTCACGCTCACCCAGCCGCGCGGAAACGTGGACCTGGGTGTGGGCTACACCGCCGTCGCGGCCACGGCCACCGTCCGCAACCCCGTTACGGACATGGGCAACCTCCCCCAAAACTTTTACCTCGGCCGCGGGGCCCCCCCGCTGCTAGACAACGCGGCCGCCGTGTACCTGCGCAACGCGGTCGTGGCGGGAAACCGGCTGGGGCCGGCCCAGCCCCTCCCGGTCTTTGGCTGCGCCCAGGTGCCGCGGCGCGCCGGCATGGACCACGGGCAGGATGCCGTGTGTGAGTTCATCGCCACCCCCGTGGCCACGGACATCAACTACTTTCGCCGGCCCTGCAACCCGCGGGGACGCGCGGCCGGCGGCGTGTACGCGGGGGACAAGGAGGGGGACGTCATAGCCCTCATGTACGACCACGGCCAGAGCGACCCGGCGCGGCCCTTCGCGGCCACGGCCAACCCGTGGGCGTCGCAGCGGTTCTCGTACGGGGACCTGCTGTACAACGGGGCCTATCACCTCAACGGGGCCTCGCCCGTCCTCAGCCCCTGCTTCAAGTTCTTCACCGCGGCCGACATCACGGCCAAACATCGCTGCCTGGAGCGTCTTATCGTGGAAACGGGATCGGCGGTATCCACGGCCACCGCTGCCAGCGACGTGCAGTTTAAGCGCCCGCCGGGGTGCCGCGAGCTCGTGGAAGACCCGTGCGGCCTGTTTCAGGAAGCCTACCCGATCACCTGCGCCAGCGACCCCGCCCTGCTACGCAGCGCCCGCGATGGGGAGGCCCACGCGCGAGAGACCCACTTTACGCAGTATCTCATCTACGACGCCTCCCCGCTAAAGGGCCTGTCTCTGTAASEQ ID NO: 121 = RS1.1Atgagtgccgaacagcgtaaaaagaaaaaaaccaccaccacgacccaaggacgtggagctgaagttgctatggcggatgaggatggaggccgcttgagagctgctgctgagactactggaggacctggatcaccggaccctgccgatggacccccccctacaccaaaccccgatcgtagaccggctgctagacctggattcggatggcatggaggacccgaggaaaacgaggacgaggcggacgacgccgctgccgacgccgacgccgatgaggctgcccctgcttctggagaggcggtagacgaacctgctgccgatggagttgttagccctaggcaattggctttgttggcgagcatggtagacgaggctgtgagaacaatcccttcccctccccctgaacgtgatggagcacaagaggaggcggctaggagtccctcaccaccccgtacaccttctatgagagcggattacggcgaggaaaacgacgacgacgacgatgatgatgacgacgatgatcgtgatgccggacgctgggttaggggacctgaaaccacttctgctgtccgtggagcataccccgatcctatggcgagtttgagccctagaccacctgccccgaggagacaccaccaccaccaccatcataggcgtagacgtgctcctagacgtcgttctgccgctagtgactcttccaaatctggctcttcttcatctgcctcttccgcttcatcttcggcctcatcgtcctcttcggcatccgcttcgagtagtgatgatgatgatgacgacgacgctgctagagcccccgcttctgctgccgaccacgctgctggcggaactttgggagccgacgacgaggaggcgggagttcctgctcgtgccccgggagctgctccgaggccttctccaccccgtgctgaacctgctccggctagaacaccggccgctactgctggtagactggagcgtagacgtgcccgtgctgctgtggctggtagagatgctactggccgcttcactgctggccgtcctagacgtgttgaactggacgccgatgctgcttctggtgctttctacgcccgttaccgtgatggttacgtgtctggtgaaccttggcctggcgctggtccacctccgcccggacgtgtactctacggtggattgggcgattctcgccctggtctgtggggcgctccg SEQ ID NO: 122 = RS1.3.1tcgagtgccgccgctgctgccgccgatttgttgttccaaaaccaatccctccgccctctgctcgccgacactgttgccgctgccgattctctggctgctccggcttctgccccacgtgaagctcgtaaacgtaaatcacccgctccggctcgtgctccccctggtggcgcccctagaccccctaaaaaatcccgtgccgatgcccctagacctgctgctgctccccccgctggtgctgctccccccgctccccctactccccccccacgcccacctcgtcccgctgccctcacacgccgtcctgctgagggacccgatccacaaggcggctggcgtagacaacctcctggcccatcccatacaccggcaccatctgccgctgctttggaggcttactgtgctcctcgtgctgtggctgaactcaccgatcatccgctgttccctgctccctggcgtcccgccctcatgttcgatcctagagctttggcttccttggccgctcgttgtgctgcccctccccctggcggtgctccggctgctttcggtcctctccgtgcctctggtccactccgccgtgccgctgcctggatgagacaagttcccgaccctgaggatgttagagttgtgatcttgtactcgcccttgcctggcgaggatttggccgctggtagagctggcggtggcccccctcctgaatggtctgctgaacgtggtggtttgtcttgcttgttggccgccctgggaaaccgtctgtgtggtcctgctactgctgcttgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaa SEQ ID NO: 123 =RS1.3.2Tgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaaggagttttgctgctctctactcgtgacttggcattcgctggagctgttgaattcctgggactcttggctggcgcttgtgataggagactcatcgtcgtaaacgctgtgagagctgccgattggcctgccgatggtcctgttgtgtctcgtcaacacgcttacttggcttgtgaagtgttgcccgctgtccaatgtgctgttcgctggcctgctgctcgtgatctgaggcgtactgttctggctagtggtcgtgttttcggacctggtgttttcgctcgtgtcgaagctgctcacgctagactgtaccccgatgccccacccctccgtttgtgtcgtggagcaaacgttcgctaccgtgtccgtactcgtttcggacccgatactctggttccaatgtcccctcgtgaataccgtcgtgctgttctgcctgccctcgatggacgtgctgccgcttctggcgctggtgacgctatggctcctggcgctccggacttctgtgaggatgaggctcactcacatcgtgcctgtgcccgctggggactgggcgctccattgaggcctgtatacgtggcactgggccgtgatgctgttagaggcggacccgctgaattgagaggccctcgtcgtgaattctgtgctagggctctgctcgaacccgatggagatgctcctcctttggtactccgtgacgacgccgatgctggtcctcccccacaaattcgctgggctagtgctgctggacgtgctggtactgtattggctgctgctggcggtggcgttgaagttgttggtactgccgctggactcgctacacctccccgccgtgaacctgtagacatggatgctgaactcgaggatgatgacgacggattgttcggagag SEQ ID NO: 124 = RS1.3tcgagtgccgccgctgctgccgccgatttgttgttccaaaaccaatccctccgccctctgctcgccgacactgttgccgctgccgattctctggctgctccggcttctgccccacgtgaagctcgtaaacgtaaatcacccgctccggctcgtgctccccctggtggcgcccctagaccccctaaaaaatcccgtgccgatgcccctagacctgctgctgctccccccgctggtgctgctccccccgctccccctactccccccccacgcccacctcgtcccgctgccctcacacgccgtcctgctgagggacccgatccacaaggcggctggcgtagacaacctcctggcccatcccatacaccggcaccatctgccgctgctttggaggcttactgtgctcctcgtgctgtggctgaactcaccgatcatccgctgttccctgctccctggcgtcccgccctcatgttcgatcctagagctttggcttccttggccgctcgttgtgctgcccctccccctggcggtgctccggctgctttcggtcctctccgtgcctctggtccactccgccgtgccgctgcctggatgagacaagttcccgaccctgaggatgttagagttgtgatcttgtactcgcccttgcctggcgaggatttggccgctggtagagctggcggtggcccccctcctgaatggtctgctgaacgtggtggtttgtcttgcttgttggccgccctgggaaaccgtctgtgtggtcctgctactgctgcttgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaaggagttttgctgctctctactcgtgacttggcattcgctggagctgttgaattcctgggactcttggctggcgcttgtgataggagactcatcgtcgtaaacgctgtgagagctgccgattggcctgccgatggtcctgttgtgtctcgtcaacacgcttacttggcttgtgaagtgttgcccgctgtccaatgtgctgttcgctggcctgctgctcgtgatctgaggcgtactgttctggctagtggtcgtgttttcggacctggtgttttcgctcgtgtcgaagctgctcacgctagactgtaccccgatgccccacccctccgtttgtgtcgtggagcaaacgttcgctaccgtgtccgtactcgtttcggacccgatactctggttccaatgtcccctcgtgaataccgtcgtgctgttctgcctgccctcgatggacgtgctgccgcttctggcgctggtgacgctatggctcctggcgctccggacttctgtgaggatgaggctcactcacatcgtgcctgtgcccgctggggactgggcgctccattgaggcctgtatacgtggcactgggccgtgatgctgttagaggcggacccgctgaattgagaggccctcgtcgtgaattctgtgctagggctctgctcgaacccgatggagatgctcctcctttggtactccgtgacgacgccgatgctggtcctcccccacaaattcgctgggctagtgctgctggacgtgctggtactgtattggctgctgctggcggtggcgttgaagttgttggtactgccgctggactcgctacacctccccgccgtgaacctgtagacatggatgctgaactcgaggatgatgacgacggattgttcggagag SEQ ID NO: 125 = RS1.4actgctggccgtcctagacgtgttgaactggacgccgatgctgcttctggtgctttctacgcccgttaccgtgatggttacgtgtctggtgaaccttggcctggcgctggtccacctccgcccggacgtgtactctacggtggattgggcgattctcgccctggtctgtggggcgctccggaggctgaggaggctagagcccgtttcgaggcttctggtgcccctgctcctgtttgggctcctgaattgggcgacgctgctcaacaatacgccctcatcacacgcttgctgtacactcccgacgccgaggctatgggatggctccaaaaccctagagttgcccctggtgatgttgctctggatcaggcttgtttccgtatctccggcgctgctcgtaactcttcttcgttcatctccggttctgtggctagagctgtgcctcacttgggatacgccatggccgctggacgtttcggctggggactggctcatgttgctgccgctgtagcaatgtctagacgctacgaccgtgctcaaaaaggattcttgctcacgtcactgaggcgtgcttacgcccctttgttggcccgtgaaaacgctgccctcactggcgcccgtacccccgatgacggtggcgacgccaaccgccacgatggtgatgatgctagaggcaaacccgctgccgctgctgctcctttgccctctgccgccgcttcccctgccgatgaacgtgctgttcctgccggttacggtgccgctggtgtgttggctgctttgggacgcttgagtgctgccccggctagtgcccccgctggtgccgatgacgatgacgatgacgatggtgctggcggaggcggtggcggtagacgtgctgaggctggacgtgttgctgttgaatgcctggctgcctgtagaggaatcttggaggctctggccgagggattcgacggagacttggcggctgtaccgggactggcgggagcgaggcctgccgctccacctcgccccggtcctgctggtgctgccgctcctcctcatgccgacgctcctagactccgtgcttggctccgtgaactccgtttcgttcgtgacgctttggttctgatgagactgagaggcgacttgagagtggctggaggatccgaggctgctgttgctgctgtccgtgctgtttctttggttgctggtgctttgggccctgctttgccgagatctccccgtttgttgtcgagtgccgccgctgctgccgccgatttgttgttccaaaaccaatccctccgccctctgctcgccgacactgttgccgctgccgattctctggctgctccggcttctgccccacgtgaagctcgtaaacgtaaatcacccgctccggctcgtgctccccctggtggcgcccctagaccccctaaaaaatcccgtgccgatgcccctagacctgctgctgctccccccgctggtgctgctccccccgctccccctactccccccccacgcccacctcgtcccgctgccctcacacgccgtcctgctgagggacccgatccacaaggcggctggcgtagacaacctcctggcccatcccatacaccggcaccatctgccgctgctttggaggcttactgtgct SEQ ID NO: 126 = RS1.5gccgctgccgattctctggctgctccggcttctgccccacgtgaagctcgtaaacgtaaatcacccgctccggctcgtgctccccctggtggcgcccctagaccccctaaaaaatcccgtgccgatgcccctagacctgctgctgctccccccgctggtgctgctccccccgctccccctactccccccccacgcccacctcgtcccgctgccctcacacgccgtcctgctgagggacccgatccacaaggcggctggcgtagacaacctcctggcccatcccatacaccggcaccatctgccgctgctttggaggcttactgtgctcctcgtgctgtggctgaactcaccgatcatccgctgttccctgctccctggcgtcccgccctcatgttcgatcctagagctttggcttccttggccgctcgttgtgctgcccctccccctggcggtgctccggctgctttcggtcctctccgtgcctctggtccactccgccgtgccgctgcctggatgagacaagttcccgaccctgaggatgttagagttgtgatcttgtactcgcccttgcctggcgaggatttggccgctggtagagctggcggtggcccccctcctgaatggtctgctgaacgtggtggtttgtcttgcttgttggccgccctgggaaaccgtctgtgtggtcctgctactgctgcttgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaaggagttttgctgctctctactcgtgacttggcattcgctggagctgttgaattcctgggactcttggctggcgcttgtgataggagactcatcgtcgtaaacgctgtgagagctgccgattggcctgccgatggtcctgttgtgtctcgtcaacacgcttacttggcttgtgaagtgttgcccgctgtccaatgtgctgttcgctggcctgctgctcgtgatctgaggcgtactgttctggctagtggtcgtgttttcggacctggtgttttcgctcgtgtcgaagctgctcacgctagactgtaccccgatgccccacccctccgtttgtgtcgtggagcaaacgttcgctaccgtgtccgtactcgtttcggacccgatactctggttccaatgtcccctcgtgaataccgtcgtgctgttctgcctgccctcgatggacgtgctgccgcttctggcgctggtgacgctatggctcctggcgctccggacttctgtgaggatgaggctcactcacatcgtgcctgtgcccgctggggactgggcgctccattgaggcctgtatacgtggcactgggccgtgatgctgttagaggcggacccgctgaattgagaggccctcgtcgtgaattctgtgctagggctctgctcgaacccgatggagatgctcctcctttggtactccgtgacgacgccgatgctggtcctcccccacaaattcgctgggctagtgctgctggacgtgctggtactgtattggctgctgctggcggtggcgttgaagttgttggtactgccgctggactcgctacacctccccgccgtgaacctgtagacatggatgctgaactcgaggatgatgacgacggattgttcggagag SEQ ID NO: 127 = RS1.6caccaccaccaccaccatcataggcgtagacgtgctcctagacgtcgttctgccgctagtgactcttccaaatctggctcttcttcatctgcctcttccgcttcatcttcggcctcatcgtcctcttcggcatccgcttcgagtagtgatgatgatgatgacgacgacgctgctagagcccccgcttctgctgccgaccacgctgctggcggaactttgggagccgacgacgaggaggcgggagttcctgctcgtgccccgggagctgctccgaggccttctccaccccgtgctgaacctgctccggctagaacaccggccgctactgctggtagactggagcgtagacgtgcccgtgctgctgtggctggtagagatgctactggccgcttcactgctggccgtcctagacgtgttgaactggacgccgatgctgcttctggtgctttctacgcccgttaccgtgatggttacgtgtctggtgaaccttggcctggcgctggtccacctccgcccggacgtgtactctacggtggattgggcgattctcgccctggtctgtggggcgctccggaggctgaggaggctagagcccgtttcgaggcttctggtgcccctgctcctgtttgggctcctgaattgggcgacgctgctcaacaatacgccctcatcacacgcttgctgtacactcccgacgccgaggctatgggatggctccaaaaccctagagttgcccctggtgatgttgctctggatcaggcttgtttccgtatctccggcgctgctcgtaactcttcttcgttcatctccggttctgtggctagagctgtgcctcacttgggatacgccatggccgctggacgtttcggctggggactggctcatgttgctgccgctgtagcaatgtctagacgctacgaccgtgctcaaaaaggattcttgctcacgtcactgaggcgtgcttacgcccctttgttggcccgtgaaaacgctgccctcactggcgcccgtacccccgatgacggtggcgacgccaaccgccacgatggtgatgatgctagaggcaaacccgctgccgctgctgctcctttgccctctgccgccgcttcccctgccgatgaacgtgctgttcctgccggttacggtgccgctggtgtgttggctgctttgggacgcttgagtgctgccccggctagtgcccccgctggtgccgatgacgatgacgatgacgatggtgctggcggaggcggtggcggtagacgtgctgaggctggacgtgttgctgttgaatgcctggctgcctgtagaggaatcttggaggctctggccgagggattcgacggagacttggcggctgtaccgggactggcgggagcgaggcctgccgctccacctcgccccggtcctgctggtgctgccgctcctcctcatgccgacgctcctagactccgtgcttggctccgtgaactccgtttcgttcgtgacgctttggttctgatgagactgagaggcgacttgagagtggctggaggatccgaggctgctgttgctgctgtccgtgctgtttctttggttgctggtgctttgggccctgctttgccgagatctccccgtttgttgtcgagtgccgccgctgctgccgccgatttgttgttccaaaaccaatccctccgccctctgctcgccgacactgttgccgctgccgattctctggctgctccggcttctgccccacgtgaagctcgtaaacgtaaatcacccgctccggctcgtgctccccctggtggcgcccctagaccccctaaaaaatcccgtgccgatgcccctagacctgctgctgctccccccgctggtgctgctccccccgctccccctactccccccccacgcccacctcgtcccgctgccctcacacgccgtcctgctgagggacccgatccacaaggcggctggcgtagacaacctcctggcccatcccatacaccggcaccatctgccgctgctttggaggcttactgtgctcctcgtgctgtggctgaactcaccgatcatccgctgttccctgctccctggcgtcccgccctcatgttcgatcctagagctttggcttccttggccgctcgttgtgctgcccctccccctggcggtgctccggctgctttcggtcctctccgtgcctctggtccactccgccgtgccgctgcctggatgagacaagttcccgaccctgaggatgttagagttgtgatcttgtactcgcccttgcctggcgaggatttggccgctggtagagctggcggtggcccccctcctgaatggtctgctgaacgtggtggtttgtcttgcttgttggccgccctgggaaaccgtctgtgtggtcctgctactgctgcttgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaaggagttttgctgctctctactcgtgacttggcattcgctggagctgttgaattcctgggactcttggctggcgcttgtgataggagactcatcgtcgtaaacgctgtgagagctgccgattggcctgccgatggtcctgttgtgtctcgtcaacacgcttacttggcttgtgaagtgttgcccgctgtccaatgtgctgttcgctggcctgctgctcgtgatctgaggcgtactgttctggctagtggtcgtgttttcggacctggtgttttcgctcgtgtcgaagctgctcacgctagactgtaccccgatgccccacccctccgtttgtgtcgtggagcaaacgttcgctaccgtgtccgtactcgtttcggacccgatactctggttccaatgtcccctcgtgaataccgtcgtgctgttctgcctgccctcgatggacgtgctgccgcttctggcgctggtgacgctatggctcctggcgctccggacttctgtgaggatgaggctcactcacatcgtgcctgtgcccgctggggactgggcgctccattgaggcctgtatacgtggcactgggccgtgatgctgttagaggcggacccgctgaattgagaggccctcgtcgtgaattctgtgctagggctctgctcgaacccgatggagatgctcctcctttggtactccgtgacgacgccgatgctggtcctcccccacaaattcgctgggctagtgctgctggacgtgctggtactgtattggctgctgctggcggtggcgttgaagttgttggtactgccgctggactcgctacacctccccgccgtgaacctgtagacatggatgctgaactcgaggatgatgacgacggattgttcggagagtaa   SEQ ID NO: 128 = RS1.7atgagtgccgaacagcgtaaaaagaaaaaaaccaccaccacgacccaaggacgtggagctgaagttgctatggcggatgaggatggaggccgcttgagagctgctgctgagactactggaggacctggatcaccggaccctgccgatggacccccccctacaccaaaccccgatcgtagaccggctgctagacctggattcggatggcatggaggacccgaggaaaacgaggacgaggcggacgacgccgctgccgacgccgacgccgatgaggctgcccctgcttctggagaggcggtagacgaacctgctgccgatggagttgttagccctaggcaattggctttgttggcgagcatggtagacgaggctgtgagaacaatcccttcccctccccctgaacgtgatggagcacaagaggaggcggctaggagtccctcaccaccccgtacaccttctatgagagcggattacggcgaggaaaacgacgacgacgacgatgatgatgacgacgatgatcgtgatgccggacgctgggttaggggacctgaaaccacttctgctgtccgtggagcataccccgatcctatggcgagtttgagccctagaccacctgccccgaggagacaccaccaccaccaccatcataggcgtagacgtgctcctagacgtcgttctgccgctagtgactcttccaaatctggctcttcttcatctgcctcttccgcttcatcttcggcctcatcgtcctcttcggcatccgcttcgagtagtgatgatgatgatgacgacgacgctgctagagcccccgcttctgctgccgaccacgctgctggcggaactttgggagccgacgacgaggaggcgggagttcctgctcgtgccccgggagctgctccgaggccttctccaccccgtgctgaacctgctccggctagaacaccggccgctactgctggtagactggagcgtagacgtgcccgtgctgctgtggctggtagagatgctactggccgcttcactgctggccgtcctagacgtgttgaactggacgccgatgctgcttctggtgctttctacgcccgttaccgtgatggttacgtgtctggtgaaccttggcctggcgctggtccacctccgcccggacgtgtactctacggtggattgggcgcccgtacccccgatgacggtggcgacgccaaccgccacgatggtgatgatgctagaggcaaacccgctgccgctgctgctcctttgccctctgccgccgcttcccctgccgatgaacgtgctgttcctgccggttacggtgccgctggtgtgttggctgctttgggacgcttgagtgctgccccggctagtgcccccgctggtgccgatgacgatgacgatgacgatggtgctggcggaggcggtggcggtagacgtgctgaggctggacgtgttgctgttgaatgcctggctgcctgtagaggaatcttggaggctctggccgagggattcgacggagacttggcggctgtaccgggactggcgggagcgaggcctgccgctccacctcgccccggtcctgctggtgctgccgctcctcctcatgccgacgctcctagactccgtgcttggctccgtgaactccgtttcgttcgtgacgctttggttctgatgagactgagaggcgacttgagagtggctggaggatccgaggctgctgttgctgctgtccgtgctgtttctttggttgctggtgctttgggccctgctttgccgagatctccccgtttgttgtcgagtgccgccgctgctgccgccgatttgttgttccaaaaccaatccctccgccctctgctcgccgacactgttgccgctgccgattctctggctgctccggcttctgccccacgtgaagctcgtaaacgtaaatcacccgctccggctcgtgctccccctggtggcgcccctagaccccctaaaaaatcccgtgccgatgcccctagacctgctgctgctccccccgctggtgctgctccccccgctccccctactccccccccacgcccacctcgtcccgctgccctcacacgccgtcctgctgagggacccgatccacaaggcggctggcgtagacaacctcctggcccatcccatacaccggcaccatctgccgctgctttggaggcttactgtgctcctcgtgctgtggctgaactcaccgatcatccgctgttccctgctccctggcgtcccgccctcatgttcgatcctagagctttggcttccttggccgctcgttgtgctgcccctccccctggcggtgctccggctgctttcggtcctctccgtgcctctggtccactccgccgtgccgctgcctggatgagacaagttcccgaccctgaggatgttagagttgtgatcttgtactcgcccttgcctggcgaggatttggccgctggtagagctggcggtggcccccctcctgaatggtctgctgaacgtggtggtttgtcttgcttgttggccgccctgggaaaccgtctgtgtggtcctgctactgctgcttgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaaggagttttgctgctctctactcgtgacttggcattcgctggagctgttgaattcctgggactcttggctggcgcttgtgataggagactcatcgtcgtaaacgctgtgagagctgccgattggcctgccgatggtcctgttgtgtctcgtcaacacgcttacttggcttgtgaagtgttgcccgctgtccaatgtgctgttcgctggcctgctgctcgtgatctgaggcgtactgttctggctagtggtcgtgttttcggacctggtgttttcgctcgtgtcgaagctgctcacgctagactgtaccccgatgccccacccctccgtttgtgtcgtggagcaaacgttcgctaccgtgtccgtactcgtttcggacccgatactctggttccaatgtcccctcgtgaataccgtcgtgctgttctgcctgccctcgatggacgtgctgccgcttctggcgctggtgacgctatggctcctggcgctccggacttctgtgaggatgaggctcactcacatcgtgcctgtgcccgctggggactgggcgctccattgaggcctgtatacgtggcactgggccgtgatgctgttagaggcggacccgctgaattgagaggccctcgtcgtgaattctgtgctagggctctgctcgaacccgatggagatgctcctcctttggtactccgtgacgacgccgatgctggtcctcccccacaaattcgctgggctagtgctgctggacgtgctggtactgtattggctgctgctggcggtggcgttgaagttgttggtactgccgctggactcgctacacctccccgccgtgaacctgtagacatggatgctgaactcgaggatgatgacgacggattgttcggagag SEQ ID NO: 129 = RS1.8atgagtgccgaacagcgtaaaaagaaaaaaaccaccaccacgacccaaggacgtggagctgaagttgctatggcggatgaggatggaggccgcttgagagctgctgctgagactactggaggacctggatcaccggaccctgccgatggacccccccctacaccaaaccccgatcgtagaccggctgctagacctggattcggatggcatggaggacccgaggaaaacgaggacgaggcggacgacgccgctgccgacgccgacgccgatgaggctgcccctgcttctggagaggcggtagacgaacctgctgccgatggagttgttagccctaggcaattggctttgttggcgagcatggtagacgaggctgtgagaacaatcccttcccctccccctgaacgtgatggagcacaagaggaggcggctaggagtccctcaccaccccgtacaccttctatgagagcggattacggcgaggaaaacgacgacgacgacgatgatgatgacgacgatgatcgtgatgccggacgctgggttaggggacctgaaaccacttctgctgtccgtggagcataccccgatcctatggcgagtttgagccctagaccacctgccccgaggagacaccaccaccaccaccatcataggcgtagacgtgctcctagacgtcgttctgccgctagtgactcttccaaatctggctcttcttcatctgcctcttccgcttcatcttcggcctcatcgtcctcttcggcatccgcttcgagtagtgatgatgatgatgacgacgacgctgctagagcccccgcttctgctgccgaccacgctgctggcggaactttgggagccgacgacgaggaggcgggagttcctgctcgtgccccgggagctgctccgaggccttctccaccccgtgctgaacctgctccggctagaacaccggccgctactgctggtagactggagcgtagacgtgcccgtgctgctgtggctggtagagatgctactggccgcttcactgctggccgtcctagacgtgttgaactggacgccgatgctgcttctggtgctttctacgcccgttaccgtgatggttacgtgtctggtgaaccttggcctggcgctggtccacctccgcccggacgtgtactctacggtggattgggcgattctcgccctggtctgtggggcgctccggaggctgaggaggctagagcccgtttcgaggcttctggtgcccctgctcctgtttgggctcctgaattgggcgacgctgctcaacaatacgccctcatcacacgcttgctgtacactcccgacgccgaggctatgggatggctccaaaaccctagagttgcccctggtgatgttgctctggatcaggcttgtttccgtatctccggcgctgctcgtaactcttcttcgttcatctccggttctgtggctagagctgtgcctcacttgggatacgccatggccgctggacgtttcggctggggactggctcatgttgctgccgctgtagcaatgtctagacgctacgaccgtgctcaaaaaggattcttgctcacgtcactgaggcgtgcttacgcccctttgttggcccgtgaaaacgctgccctcactggcgcccgtacccccgatgacggtggcgacgccaaccgccacgatggtgatgatgctagaggcaaacccgctgccgctgctgctcctttgccctctgccgccgcttcccctgccgatgaacgtgctgttcctgccggttacggtgccgctggtgtgttggctgctttgggacgcttgagtgctgccccggctagtgcccccgctggtgccgatgacgatgacgatgacgatggtgctggcggaggcggtggcggtagacgtgctgaggctggacgtgttgctgttgaatgcctggctgcctgtagaggaatcttggaggctctggccgagggattcgacggagacttggcggctgtaccgggactggcgggagcgaggcctgccgctccacctcgccccggtcctgctggtgctgccgctcctcctcatgccgacgctcctagactccgtgcttggctccgtgaactccgtttcgttcgtgacgctttggttctgatgagactgagaggcgacttgagagtggctggaggatccgaggctgctgttgctgctgtccgtgctgtttctttggttgctggtgctttgggccctgctttgccgagatctccccgtttgttgtcgagtgccgccgctgctgccgccgatttgttgttccaaaaccaatccctccgccctctgctcgccgacactgttgccgctgccgattctctggctgctccggcttctacaccggcaccatctgccgctgctttggaggcttactgtgctcctcgtgctgtggctgaactcaccgatcatccgctgttccctgctccctggcgtcccgccctcatgttcgatcctagagctttggcttccttggccgctcgttgtgctgcccctccccctggcggtgctccggctgctttcggtcctctccgtgcctctggtccactccgccgtgccgctgcctggatgagacaagttcccgaccctgaggatgttagagttgtgatcttgtactcgcccttgcctggcgaggatttggccgctggtagagctggcggtggcccccctcctgaatggtctgctgaacgtggtggtttgtcttgcttgttggccgccctgggaaaccgtctgtgtggtcctgctactgctgcttgggctggaaactggactggcgctcccgatgtttctgctctcggtgctcaaggagttttgctgctctctactcgtgacttggcattcgctggagctgttgaattcctgggactcttggctggcgcttgtgataggagactcatcgtcgtaaacgctgtgagagctgccgattggcctgccgatggtcctgttgtgtctcgtcaacacgcttacttggcttgtgaagtgttgcccgctgtccaatgtgctgttcgctggcctgctgctcgtgatctgaggcgtactgttctggctagtggtcgtgttttcggacctggtgttttcgctcgtgtcgaagctgctcacgctagactgtaccccgatgccccacccctccgtttgtgtcgtggagcaaacgttcgctaccgtgtccgtactcgtttcggacccgatactctggttccaatgtcccctcgtgaataccgtcgtgctgttctgcctgccctcgatggacgtgctgccgcttctggcgctggtgacgctatggctcctggcgctccggacttctgtgaggatgaggctcactcacatcgtgcctgtgcccgctggggactgggcgctccattgaggcctgtatacgtggcactgggccgtgatgctgttagaggcggacccgctgaattgagaggccctcgtcgtgaattctgtgctagggctctgctcgaacccgatggagatgctcctcctttggtactccgtgacgacgccgatgctggtcctcccccacaaattcgctgggctagtgctgctggacgtgctggtactgtattggctgctgctggcggtggcgttgaagttgttggtactgccgctggactcgctacacctccccgccgtgaacctgtagacatggatgctgaactcgaggatgatgacgacggattgttcggagag SEQ ID NO: 130 = His tag HHHHHH SEQ ID NO: 131 = Tag MSYYHHHHHHSEQ ID NO: 132 = Secretion Signal MKFLVNVALVFMVVYISYIYA SEQ ID NO: 133 =UL49.5ATGTCGTACTACCATCACCATCACCATCACATGACGGGGAAACCCGCAAGACTGGGCCGCTGGGTGGTGCTGTTGTTCGTCGCGCTCGTCGCGGGCGTGCCCGGGGAGCCGCCGAACGCGGCAGGCGCACGCGGCGTTATCGGGGACGCGCAATGCCGGGGCGACAGCGCCGGTGTGGTGTCCGTCCCGGGGGTCCTGGTGCCCTTTTATCTAGGCATGACCTCGATGGGCGTATGTATGATCGCGCACGTGTATCAGATATGCCAGCGGGCACTGGCCGCCGGGTCAGCCTGASEQ ID NO: 134 = UL10ATGGGACGCCGGGCCCCCAGGGGATCCCCCGAGGCCGCGCCGGGCGCCGACGTCGCGCCCGGGGCGCGGGCGGCGTGGTGGGTCTGGTGTGTGCAGGTGGCGACGTTCATCGTCTCGGCCATCTGCGTCGTGGGGCTCCTGGTGCTGGCCTCTGTGTTCCGGGACAGGTTTCCCTGCCTTTACGCCCCCGCGACCTCTTATGCGAAGGCGAACGCCACGGTCGAGGTGCGCGGGGGTGTAGCCGTCCCCCTCCGGTTGGACACGCAGAGCCTGCTGGCCACGTACGCAATTACGTCTACGCTGTTGCTGGCGGCGGCCGTGTACGCCGCGGTGGGCGCGGTGACCTCGCGCTACGAGCGCGCGCTGGATGCGGCCCGTCGCCTGGCGGCGGCCCGTATGGCGATGCCACACGCCACGCTAATCGCCGGAAACGTCTGCGCGTGGCTGTTGCAGATCACAGTCCTGCTGCTGGCCCACCGCATCAGCCAGCTGGCCCACCTTATCTACGTCCTGCACTTTGCGTGCCTCGTGTATCTCGCGGCCCATTTTTGCACCAGGGGGGTCCTGAGCGGGACGTACCTGCGTCAGGTTCACGGCCTGATTGACCCGGCGCCGACGCACCATCGTATCGTCGGTCCGGTGCGGGCAGTAATGACAAACGCCTTATTACTGGGCACCCTCCTGTGCACGGCCGCCGCCGCGGTCTCGTTGAACACGATCGCCGCCCTGAACTTCAACTTTTCCGCCCCGAGCATGCTCATCTGCCTGACGACGCTGTTCGCCCTGCTTGTCGTGTCGCTGTTGTTGGTGGTCGAGGGGGTGCTGTGTCACTACGTGCGCGTGTTGGTGGGCCCCCACCTCGGGGCCATCGCCGCCACCGGCATCGTCGGCCTGGCCTGCGAGCACTACCACACCGGTGGTTACTACGTGGTGGAGCAGCAGTGGCCGGGGGCCCAGACGGGAGTCCGCGTCGCCCTGGCGCTCGTCGCCGCCTTTGCCCTCGCCATGGCCGTGCTTCGGTGCACGCGCGCCTACCTGTATCACCGGCGACACCACACTAAATTTTTCGTGCGCATGCGCGACACCCGGCACCGCGCCCATTCGGCGCTTCGACGCGTACGCAGCTCCATGCGCGGTTCTAGGCGTGGCGGGCCGCCCGGAGACCCGGGCTACGCGGAAACCCCCTACGCGAGCGTGTCCCACCACGCCGAGATCGACCGGTATGGGGATTCCGACGGGGACCCGATCTACGACGAAGTGGCCCCCGACCACGAGGCCGAGCTCTACGCCCGAGTGCAACGCCCCGGGCCTGTGCCCGACGCCGAGCCCATTTACGACACCGTGGAGGGGTATGCGCCAAGGTCCGCGGGGGAGCCGGTGTACAGCACCGTTCGGCGATGGTAG

The invention claimed is:
 1. An immunogenic composition comprising a pharmaceutically-acceptable carrier, an immunostimulatory amount of an adjuvant and a particulate composition comprising a polypeptide comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 2, wherein the polypeptide does not comprise a full length ICP4 polypeptide.
 2. A method of treating a subject suffering from HSV-2 infection, comprising administering to the subject an effective amount of the immunogenic composition according to claim
 1. 3. The immunogenic composition of claim 1, wherein upon administration to a subject infected with HSV-2, the immunogenic composition inhibits HSV-2 symptoms.
 4. The immunogenic composition of claim 1, wherein upon administration to a subject infected with HSV-2, the immunogenic composition reduces the number of herpetic lesions.
 5. The immunogenic composition of claim 1, wherein upon administration to a subject infected with HSV-2, the immunogenic composition reduces the number of days a subject experiences herpetic lesions.
 6. A method of treating a subject suffering from HSV-2 infection, comprising: administering to the subject an initial dose of an effective amount of an immunogenic composition; and administering to the subject one or more additional doses of the immunogenic composition at a specified time interval after receiving the initial dose; wherein the immunogenic composition comprises a formulation comprising a pharmaceutically-acceptable carrier, an immunostimulatory amount of an adjuvant and a particulate composition comprising a polypeptide comprising an amino acid sequence having at least 90% identity to SEQ ID NO:
 2. 7. The method of claim 6, comprising administering one or more additional doses of the immunogenic composition 1 month, 2 months, 4 months, 6 months or 12 months after receiving the initial dose.
 8. The method of claim 6, wherein the one or more additional doses comprises a first and a second additional dose.
 9. The method of claim 6, wherein the method reduces the number of herpetic lesions at the onset of HSV-2 infection.
 10. The method of claim 6, wherein the one or more additional doses are administered in a dose escalation manner, such that at least one successive administration of the immunogenic composition contains a higher concentration of the immunogenic composition than at least one previous dose.
 11. The method of claim 6, wherein the one or more additional doses are administered in a manner such that at least one successive administration of the immunogenic composition contains a lower concentration of the immunogenic composition than at least one previous dose.
 12. The immunogenic composition of claim 1, wherein upon administration to a subject infected with HSV-2, the immunogenic composition activates a T cell response to one or more HSV-2 antigens in the subject.
 13. The immunogenic composition of claim 1, wherein upon administration to a subject infected with HSV-2, the immunogenic composition increases a T cell response to one or more HSV-2 antigens in a subject.
 14. The immunogenic composition of claim 1, wherein upon administration to a subject infected with HSV-2, the subject exhibits a delayed onset of symptoms or reduced severity of HSV-2 symptoms.
 15. The method of claim 2, wherein administering the immunogenic composition comprises intravenous, intraperitoneal, intramuscular, intradermal, subcutaneous, transdermal, subdermal, intracranial, intranasal, mucosal, anal, vaginal, oral, sublingual, buccal, inhaled, or topical administration.
 16. The method of claim 15, wherein administering the immunogenic composition comprises subcutaneous, intranasal, or mucosal administration.
 17. The immunogenic composition of claim 1, wherein the particulate composition further comprises an HSV gD2 polypeptide.
 18. The immunogenic composition of claim 1, wherein the particulate composition further comprises an HSV gD2 polypeptide lacking a transmembrane and/or lacking a cytoplasmic domain.
 19. The immunogenic composition of claim 1, wherein the particulate composition further comprises a second polypeptide consisting of the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4 or SEQ ID NO:
 5. 20. The immunogenic composition of claim 1, wherein the polypeptide is lacking 1-20 amino acids from the N-terminus, C-terminus, or both.
 21. The immunogenic composition of claim 1, wherein the polypeptide consists of an amino acid sequence having at least 90% identity to SEQ ID NO:2. 